P2x3 and/or p2x2/3 receptor antagonist, pharmaceutical composition comprising same, and use thereof

ABSTRACT

A P2X3 and/or P2X2/3 receptor antagonist of formula (I), a pharmaceutical composition comprising the same, and a use thereof in preparing a drug for preventing or treating a disease mediated by the P2X3 and/or P2X2/3 receptor antagonist.

FIELD OF THE INVENTION

The present invention relates to a P2X3 and/or P2X2/3 receptorantagonist, a pharmaceutical composition comprising the same, and usethereof for the prophylaxis or treatment of a disease mediated by theP2X3 and/or P2X2/3 receptor antagonist.

BACKGROUND OF THE INVENTION

Purine compounds, acting via cell surface purinoceptors, have beenimplicated as having a variety of physiological and pathological roles.ATP, and to a lesser extent, adenosine, can stimulate sensory nerveendings resulting in intense pain and a pronounced increase in sensorynerve discharge. ATP receptors have been classified into two majorfamilies, the P2Y- and P2X-purinoreceptors, on the basis of themolecular structure, transduction mechanisms, and pharmacologicalcharacterization. The P2Y-purinoceptors are G-protein coupled receptors,while the P2X-purinoceptors are a family of ATP-gated cation channels.Purinoceptors, in particular, P2X receptors, can form homomultimers orheteromultimers. To date, cDNAs for multiple P2X receptor subtypes(including six homologous receptors: P2X1, P2X2, P2X3, P2X4, P2X5 andP2X7; and three heterologous receptors: P2X2/3, P2X4/6 and P2X1/5) havebeen cloned. The structure and chromosomal mapping of mouse genomic P2X3receptor subunits have also been reported.

Studies have shown that P2X3 and/or P2X2/3 receptor antagonists can beused to treat diseases such as pain, etc. The present invention providescompounds as P2X receptor modulators, particularly P2X3 and/or P2X2/3receptor antagonists.

SUMMARY OF THE INVENTION

The present invention provides a compound for use as a P2X receptormodulator (particularly P2X3 and/or P2X2/3 receptor antagonist), iteffectively antagonize the P2X receptor (particularly P2X3 and/or P2X2/3receptor), and has better physicochemical properties (e.g., solubility,physical and/or chemical stability), improved pharmacokinetic properties(e.g., improved bioavailability, proper half-life and duration ofaction), improved safety (low toxicity and/or less side effects, widetherapeutic window), and the like.

An aspect of the present invention provides a compound or apharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein the compound has the structure of Formula (I):

L is selected from the group consisting of C(═O), CRR′, NR, O, S, S═Oand S(═O)₂;

V¹ is selected from the group consisting of N,

N—O and NR;

V² is selected from the group consisting of CR⁶ and C(═O);

represents either a single bond or a double bond, provided that when

is a single bond, V¹ is NR and V² is C(═O);

R and R′ are each independently selected from the group consisting of H,halogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated or partiallyunsaturated C₃₋₁₀ cyclic hydrocarbyl, saturated or partially unsaturated3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryland C₆₋₁₂ aralkyl, and at most 2 ring members in the cyclic hydrocarbyland heterocyclyl are C(═O);

R¹, R², R³ and R⁶ are each independently selected from the groupconsisting of H, halogen, —CN, —NH₂, —OH, —SH, —Se—R, —Si(R)₃, C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated or partially unsaturatedC₃₋₁₀ cyclic hydrocarbyl, saturated or partially unsaturated 3- to10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl,C₆₋₁₂ aralkyl, C₁₋₆ haloalkyl, —C(═O)R^(a), —OC(═O)R^(a), —C(═O)OR^(a),—OR^(a), —SR^(a), —S(═O)R^(a), —S(═O)₂R^(a), —S(═O)₂NR^(a)R^(b),—S(═O)(═NR)R^(a), —NR^(a)R^(b), —C(═O)NR^(a)R^(b), —C(═S)NR^(a)R^(b),—C(═NR)NR^(a)R^(b), —NR^(a)—C(═O)R^(b), —NR^(a)—C(═O)OR^(b),—NR^(a)—S(═O)₂—R^(b), —NR^(a)—C(═O)—NR^(a)R^(b), —C₁₋₆alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a), —C₁₋₆ alkylene-C(═O)R,—C₁₋₆ alkenylene-OR^(a), —O—C₁₋₆ alkylene-NR^(a)R^(b) and—P(═O)R^(a)R^(b);

R⁴ and R⁵ are each independently selected from the group consisting ofH, —C(═O)OR^(a), —NR^(a)R^(b), —NR^(a)—C(═O)R^(b), —NR^(a)—C(═)OR, —C₁₋₆alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a), —C₁₋₆ alkylene-O—C₁₋₆alkylene-OR^(a), C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,saturated or partially unsaturated C₃₋₁₀ cyclic hydrocarbyl, saturatedor partially unsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5-to 14-membered heteroaryl and C₆₋₁₂ aralkyl;

-   -   alternatively, R¹ and R⁴ together form —NH—(C₁₋₆        alkylene)-L-(C₁₋₆ alkylene)-, preferably —NHCH₂CH₂—O—CH₂CH₂—;    -   the above alkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbyl,        heterocyclyl, aryl, heteroaryl and aralkyl, at each occurrence,        are each optionally substituted with one or more substituents        independently selected from the group consisting of halogen,        hydroxyl, oxo, amino, cyano, nitro, —Si(R)₃, C₁₋₆ alkyl,        saturated or partially unsaturated C₃₋₆ cyclic hydrocarbyl,        saturated or partially unsaturated 3- to 10-membered        heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl, C₆₋₁₂        aralkyl, —C(═O)R^(a), —OC(═O)R^(a), —C(═O)OR^(a), —OR^(a),        —SR^(a), —S(═O)R^(a), —S(═O)₂R^(a), —S(═O)₂NR^(a)R^(b),        —NR^(a)R^(b), —C(═O)NR^(a)R^(b), —NR^(a)—C(═O)R^(b),        —NR^(a)—C(═O)OR^(b), —NR^(a)—S(═O)₂—R^(b),        —NR^(a)—C(═O)—NR^(a)R^(b), —C₁₋₆ alkylene-NR^(a)R^(b), —C₁₋₆        alkylene-OR^(a), —C₁₋₆ alkenylene-OR^(a) and —O—C₁₋₆        alkylene-NR^(a)R^(b), the alkyl, cyclic hydrocarbyl,        heterocyclyl, aryl, heteroaryl and aralkyl are further        optionally substituted with one or more substituents        independently selected from the group consisting of halogen,        hydroxyl, oxo, amino, cyano, nitro, —NR^(a)R^(b), C₁₋₆ alkyl,        —O—C₁₋₆ alkyl, saturated or partially unsaturated C₃₋₆ cyclic        hydrocarbyl, saturated or partially unsaturated 3- to        10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered        heteroaryl and C₆₋₁₂ aralkyl; and

R^(a) and R^(b), at each occurrence, are each independently selectedfrom the group consisting of H, —OH, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆alkynyl, saturated or partially unsaturated C₃₋₁₀ cyclic hydrocarbyl,saturated or partially unsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀aryl, 5- to 14-membered heteroaryl and C₆₋₁₂ aralkyl; alternatively,R^(a) and R^(b) together with the atom to which they are attached form a3- to 12-membered heterocycle or heteroaromatic ring, the above groupsare further optionally substituted with one or more substituentsindependently selected from the group consisting of halogen, hydroxyl,oxo, amino, cyano, nitro, C₁₋₆ alkyl, —O—C₁₋₆ alkyl, saturated orpartially unsaturated C₃₋₆ cyclic hydrocarbyl, saturated or partiallyunsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to14-membered heteroaryl and C₆₋₁₂ aralkyl.

Another aspect of the present invention provides a pharmaceuticalcomposition comprising a prophylactically or therapeutically effectiveamount of the compound of the present invention or a pharmaceuticallyacceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide,isotopically labeled compound, metabolite or prodrug thereof, and one ormore pharmaceutically acceptable carriers, and the pharmaceuticalcomposition is preferably in the form of a solid, semi-solid, liquid, orgas preparation.

Another aspect of the present invention provides use of the compound ofthe present invention or a pharmaceutically acceptable salt, ester,stereoisomer, polymorph, solvate, N-oxide, isotopically labeledcompound, metabolite or prodrug thereof or the pharmaceuticalcomposition of the present invention in the manufacture of a medicamentfor the treatment of a disease mediated by the P2X3 and/or P2X2/3receptor antagonist.

Another aspect of the present invention provides the compound of thepresent invention or a pharmaceutically acceptable salt, ester,stereoisomer, polymorph, solvate, N-oxide, isotopically labeledcompound, metabolite or prodrug thereof or the pharmaceuticalcomposition of the present invention for use in the treatment of adisease mediated by the P2X3 and/or P2X2/3 receptor antagonist.

Another aspect of the present invention provides a method for theprophylaxis or the treatment of a disease mediated by the P2X3 and/orP2X2/3 receptor antagonist, wherein the method comprises administeringto a subject in need thereof an effective amount of the compound of thepresent invention or a pharmaceutically acceptable salt, ester,stereoisomer, polymorph, solvate, N-oxide, isotopically labeledcompound, metabolite or prodrug thereof or the pharmaceuticalcomposition of the present invention.

DETAILED DESCRIPTION OF THE INVENTION Definition

Unless otherwise defined in the context, all technical and scientificterms used herein are intended to have the same meaning as commonlyunderstood by a person skilled in the art. References to techniquesemployed herein are intended to refer to the techniques as commonlyunderstood in the art, including variations on those techniques orsubstitutions of equivalent techniques which would be apparent to aperson skilled in the art. While it is believed that the following termswill be readily understood by a person skilled in the art, the followingdefinitions are nevertheless put forth to better illustrate the presentinvention.

The terms “contain”, “include”, “comprise”, “have”, or “relate to”, aswell as other variations used herein are inclusive or open-ended, and donot exclude additional, unrecited elements or method steps.

As used herein, the term “alkylene” refers to a saturated divalenthydrocarbyl, preferably refers to a saturated divalent hydrocarbylhaving 1, 2, 3, 4, 5 or 6 carbon atoms, e.g., methylene, ethylene,propylene or butylene.

As used herein, the term “alkyl” is defined as a linear or branchedsaturated aliphatic hydrocarbon. In some embodiments, alkyl has 1-12,e.g., 1-6, carbon atoms. For example, as used herein, the term “C₁₋₆alkyl” refers to a linear or branched group having 1-6 carbon atoms(such as methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,sec-butyl, tert-butyl, n-pentyl, isopentyl, neopentyl, or n-hexyl),which is optionally substituted with one or more (e.g., 1 to 3) suitablesubstituents such as halogen (in which case the group may be referred toas “haloalkyl”) (e.g., CH₂F, CHF₂, CF₃, CCl₃, C₂F₅, C₂Cl₅, CH₂CF₃, CH₂Clor —CH₂CH₂CF₃ etc.). The term “C₁₋₄ alkyl” refers to a linear orbranched aliphatic hydrocarbon chain having 1-4 carbon atoms (i.e.,methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl ortert-butyl).

As used herein, the term “alkenyl” refers to a linear or branchedmonovalent hydrocarbyl having a double bond and 2-6 carbon atoms (“C₂₋₆alkenyl”). The alkenyl is e.g., vinyl, 1-propenyl, 2-propenyl,2-butenyl, 3-butenyl, 2-pentenyl, 3-pentenyl, 4-pentenyl, 2-hexenyl,3-hexenyl, 4-hexenyl, 5-hexenyl, 2-methyl-2-propenyl and4-methyl-3-pentenyl. When the compound of the present invention containsan alkenylene group, the compound may exist as the pure E (entgegen)form, the pure Z (zusammen) form, or any mixture thereof.

As used herein, the term “alkynyl” refers to a monovalent hydrocarbylcontaining one or more triple bond, and preferably having 2, 3, 4, 5 or6 carbon atoms, e.g., ethynyl or propynyl.

As used herein, the term “cycloalkyl” refers to a saturated monocyclicor polycyclic (e.g., bicyclic) hydrocarbon ring (e.g., monocyclic, suchas cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl,cyclooctyl, or cyclononyl, or bicyclic, including spiro, fused orbridged cyclic system (such as bicyclo[1.1.1]pentyl,bicyclo[2.2.1]heptyl, bicyclo[3.2.1]octyl or bicyclo[5.2.0]nonyl, ordecahydronaphthalene etc.)), which is optionally substituted with one ormore (e.g., 1 to 3) suitable substituents. The cycloalkyl has 3 to 15carbon atoms. For example, the term “C₃₋₆ cycloalkyl” refers to asaturated monocyclic or polycyclic (e.g., bicyclic) hydrocarbon ringhaving 3 to 6 ring forming carbon atoms (e.g., cyclopropyl, cyclobutyl,cyclopentyl, or cyclohexyl), which is optionally substituted with one ormore (e.g., 1 to 3) suitable substituents, e.g., methyl substitutedcyclopropyl.

As used herein, the terms “cyclic hydrocarbylene”, “cyclic hydrocarbyl”and “hydrocarbon ring” refer to a saturated (i.e., “cycloalkylene” and“cycloalkyl”) or unsaturated (i.e., having one or more double and/ortriple bonds in the ring) monocyclic or polycyclic hydrocarbon ringhaving e.g., 3-10 (suitably having 3-8, and more suitably having 3-6)ring carbon atoms, including but not limited to cyclopropyl(ene) (ring),cyclobutyl(ene) (ring), cyclopentyl(ene) (ring), cyclohexyl(ene) (ring),cycloheptyl(ene) (ring), cyclooctyl(ene) (ring), cyclononyl(ene) (ring),cyclohexenyl(ene) (ring), and the like.

As used herein, the terms “heterocyclyl”, “heterocyclylene” and“heterocycle” refer to a saturated (i.e., heterocycloalkyl) or partiallyunsaturated (i.e., having one or more double and/or triple bonds in thering) cyclic group having e.g. 3-10 (suitably having 3-8, and moresuitably having 3-6) ring atoms, wherein at least one ring atom is aheteroatom selected from the group consisting of N, O and S, and theremaining ring atoms are C. For example, “3- to 10-memberedheterocyclyl(ene)” of “3- to 10-membered heterocycle” refers tosaturated or partially unsaturated heterocyclyl(ene) or heterocyclehaving 2-9 (e.g., 2, 3, 4, 5, 6, 7, 8 or 9) ring carbon atoms and one ormore (e.g., 1, 2, 3, or 4) heteroatoms independently selected from thegroup consisting of N, O and S. Examples of heterocyclylene,heterocyclyl and heterocycle include, but are not limited tooxiranyl(ene), aziridinyl(ene), azetidinyl(ene), oxetanyl(ene),tetrahydrofuranyl(ene), dioxolinyl(ene), pyrrolidinyl(ene),pyrrolidonyl(ene), imidazolidinyl(ene), pyrazolidinyl(ene),pyrrolinyl(ene), tetrahydropyranyl(ene), piperidinyl(ene),morpholinyl(ene), dithianyl(ene), thiomorpholinyl(ene), piperazinyl(ene)or trithianyl(ene). Said group also encompasses a bicyclic system,including a spiro, fused, or bridged system (e.g.,8-azaspiro[4.5]decane, 3,9-diazaspiro[5.5]undecane,2-azabicyclo[2.2.2]octane, etc.). Heterocyclylene, heterocyclyl andheterocycle may optionally be substituted with one or more (e.g. 1, 2, 3or 4) suitable substituents.

As used herein, the terms “aryl(ene)” and “aromatic ring” refer to anall-carbon monocyclic or fused-ring polycyclic aromatic group having aconjugated 7E electron system. For example, as used herein, the terms“C₆₋₁₀ aryl(ene)” and “C₆₋₁₀ aromatic ring” refer to an aromatic groupcontaining 6 to 10 carbon atoms, such as phenyl(ene) (benzene ring) ornaphthyl(ene) (naphthalene ring). Aryl(ene) or aromatic ring isoptionally substituted with one or more (such as 1 to 3) suitablesubstituents (e.g., halogen, —OH, —CN, —NO₂, and C₁₋₆ alkyl, etc.).

As used herein, the terms “heteroaryl(ene)” and “heteroaromatic ring”refer to a monocyclic, bicyclic or tricyclic aromatic ring system having5, 6, 8, 9, 10, 11, 12, 13 or 14 ring atoms, particularly 1 or 2 or 3 or4 or 5 or 6 or 9 or 10 carbon atoms, and containing at least oneheteroatom (such as O, N, or S), which can be same to different.Moreover, in each case, it can be benzo-fused. In particular,“heteroaryl(ene)” or “heteroaromatic ring” is selected from the groupconsisting of thienyl(ene), furyl(ene), pyrrolyl(ene), oxazolyl(ene),thiazolyl(ene), imidazolyl(ene), pyrazolyl(ene), isoxazolyl(ene),isothiazolyl(ene), oxadiazolyl(ene), triazolyl(ene), thiadiazolyl(ene)etc., and benzo derivatives thereof; or pyridinyl(ene),pyridazinyl(ene), pyrimidinyl(ene), pyrazinyl(ene), triazinyl(ene),etc., and benzo derivatives thereof.

As used herein, the term “aralkyl” preferably means aryl or heteroarylsubstituted alkyl, wherein aryl, heteroaryl and alkyl are as definedherein. Normally, the aryl group may have 6-14 carbon atoms, theheteroaryl group may have 5-14 ring atoms, and the alkyl group may have1-6 carbon atoms. Exemplary aralkyl group includes, but is not limitedto, benzyl, phenylethyl, phenylpropyl, phenylbutyl.

As used herein, the term “halo” or “halogen” are defined to include F,Cl, Br, or I.

As used herein, the term “nitrogen containing heterocycle” refers to asaturated or unsaturated monocyclic or bicyclic group having 2, 3, 4, 5,6, 7, 8, 9, 10, 11, 12 or 13 carbon atoms and at least one nitrogen atomin the ring, which may further optionally comprise one or more (e.g.,one, two, three or four) ring members selected from the group consistingof N, O, C═O, S, S═O and S(═O)₂. The nitrogen containing heterocycle isattached to the rest of the molecule through the nitrogen atom and anyother ring atom in said nitrogen containing heterocycle. The nitrogencontaining heterocycle is optionally benzo-fused, and is preferablyattached to the rest of the molecule through the nitrogen atom in saidnitrogen containing heterocycle and any carbon atom in the fused benzenering.

The term “substituted” means that one or more (e.g., one, two, three, orfour) hydrogens on the designated atom is replaced with a selection fromthe indicated group, provided that the designated atom's normal valencyunder the existing circumstances is not exceeded, and that thesubstitution results in a stable compound. Combinations of substituentsand/or variables are permissible only if such combinations result instable compounds.

If a substituent is described as being “optionally substituted”, thesubstituent may be either (1) not substituted, or (2) substituted. If acarbon of a substituent is described as being optionally substitutedwith one or more of a list of substituents, one or more of the hydrogenson the carbon (to the extent there are any) may separately and/ortogether be replaced with an independently selected optionalsubstituent. If a nitrogen of a substituent is described as beingoptionally substituted with one or more from a list of substituents, oneor more of the hydrogens on the nitrogen (to the extent there are any)may each be replaced with an independently selected optionalsubstituent.

If substituents are described as being “independently selected” from agroup, each substituent is selected independent of the other(s). Eachsubstituent therefore may be identical to or different from the othersubstituent(s).

As used herein, the term “one or more” means one or more than one (e.g.,2, 3, 4, 5 or 10) as reasonable.

As used herein, unless specified, the point of attachment of asubstituent can be from any suitable position of the substituent.

When a bond to a substituent is shown to cross a bond connecting twoatoms in a ring, then such substituent may be bonded to any of thering-forming atoms in that ring that are substitutable.

The present invention also includes all pharmaceutically acceptableisotopically labeled compounds, which are identical to those of thepresent invention except that one or more atoms are replaced by an atomhaving the same atomic number, but an atomic mass or mass numberdifferent from the atomic mass or mass number which predominates innature. Examples of isotopes suitable for inclusion in the compound ofthe present invention include, but are not limited to, isotopes ofhydrogen, such as ²H, ³H; carbon, such as ¹¹C, ¹³C, and ¹⁴C; chlorine,such as ³⁶Cl; fluorine, such as ¹⁸F; iodine, such as ¹²³I and ¹²⁵I;nitrogen, such as ¹³N and ¹⁵N; oxygen, such as ¹⁵O, ¹⁷O, and ¹⁸O;phosphorus, such as ³²P; and sulfur, such as ³¹S. Certain isotopicallylabeled compounds of the present invention, for example thoseincorporating a radioactive isotope, are useful in drug and/or substratetissue distribution studies (e.g., assays). The radioactive isotopestritium, i.e., ³H, and carbon-14, i.e., ¹⁴C, are particularly useful forthis purpose in view of their ease of incorporation and ready means ofdetection. Substitution with positron-emitting isotopes, such as ¹¹C,¹⁸F, ¹⁵O and ¹³N, can be useful in positron emission tomography (PET)studies for examining substrate receptor occupancy. Isotopically labeledcompounds of the present invention can generally be prepared byprocesses analogous to those described in the accompanying Schemesand/or in the Examples and Preparations, by using an appropriateisotopically labeled reagent in place of the non-labeled reagentpreviously employed. Pharmaceutically acceptable solvates in accordancewith the invention include those wherein the solvent of crystallizationmay be isotopically substituted, e.g., D₂O, acetone-d₆, or DMSO-d₆.

The term “stereoisomer” refers to isomers with at least one asymmetriccenter. A compound having one or more (e.g., one, two, three or four)asymmetric centers can give rise to a racemic mixture, singleenantiomer, diastereomer mixture and individual diastereomer. Certainindividual molecules may exist as geometric isomers (cis/trans).Similarly, the compound of the present invention may exist as a mixtureof two or more structurally different forms in rapid equilibrium(generally referred to as tautomer). Typical examples of a tautomerinclude a keto-enol tautomer, phenol-keto tautomer, nitroso-oximetautomer, imine-enamine tautomer and the like. It is to be understoodthat all such isomers and mixtures thereof in any proportion (such as60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, and 99%) areencompassed within the scope of the present invention.

The chemical bonds of the compound of the present invention may bedepicted herein using a solid line (

), a solid wedge (

), or a dotted wedge (

). The use of a solid line to depict bonds to asymmetric carbon atoms ismeant to indicate that all possible stereoisomers (e.g., specificenantiomers, racemic mixtures, etc.) at that carbon atom are included.The use of either a solid or dotted wedge to depict bonds to asymmetriccarbon atoms is meant to indicate that the stereoisomer shown ispresent. When present in racemic compounds, solid and dotted wedges areused to define relative stereochemistry, rather than absolutestereochemistry. Unless stated otherwise, it is intended that thecompound of the present invention can exist as stereoisomers, whichinclude cis and trans isomers, optical isomers such as R and Senantiomers, diastereomers, geometric isomers, rotational isomers,conformational isomers, atropisomers, and mixtures thereof. The compoundof the present invention may exhibit more than one type of isomerism,and consist of mixtures thereof (such as racemates and diastereomericpairs).

The present invention includes all possible crystalline forms orpolymorphs of the compound of the present invention, either as a singlepolymorph, or as a mixture of more than one polymorphs, in any ratio.

It also should be understood that, certain compounds of the presentinvention can be used for the treatment in a free from, or whereappropriate, in a form of a pharmaceutically acceptable derivative. Inthe present invention, the pharmaceutically acceptable derivativeincludes, but is not limited to a pharmaceutically acceptable salt,ester, solvate, N-oxide, metabolite or prodrug, which can directly orindirectly provide the compound of the present invention or a metaboliteor residue thereof after being administered to a patient in needthereof. Therefore, “the compound of the present invention” mentionedherein also means to encompass various derivative forms of the compoundas mentioned above.

A pharmaceutically acceptable salt of the compound of the presentinvention includes an acid addition salt and a base addition saltthereof.

A suitable acid addition salt is formed from an acid which forms apharmaceutically acceptable salt. Specific examples include acetate,adipate, aspartate, benzoate, besylate, bicarbonate/carbonate,bisulfate/sulfate, borate, camphorsulfonate, citrate, cyclamate,edisylate, esylate, formate, fumarate, gluceptate, gluconate,glucuronate, hexafluorophosphate, hibenzate, hydrochloride/chloride,hydrobromide/bromide, hydroiodide/iodide, isethionate, lactate, malate,maleate, malonate, mesylate, methylsulfate, naphthylate, 2-napsylate,nicotinate, nitrate, orotate, oxalate, palmitate, pamoate,phosphate/hydrogen phosphate/dihydrogen phosphate, pyroglutamate,saccharate, stearate, succinate, tannate, tartrate, tosylate,trifluoroacetate and xinofoate salts.

A suitable base addition salt is formed from a base which forms apharmaceutically acceptable salt. Specific examples include aluminum,arginine, benzathine, calcium, choline, diethylamine, diolamine,glycine, lysine, magnesium, meglumine, olamine, potassium, sodium,tromethamine and zinc salts.

For a review on suitable salts, see “Handbook of Pharmaceutical Salts:Properties, Selection, and Use” by Stahl and Wermuth (Wiley-VCH, 2002).The method for preparing a pharmaceutically acceptable salt of thecompound of the present invention is known to a person skilled in theart.

As used herein, the term “ester” refers to those derived from thecompounds of the various formulae in the present application, whichinclude physiologically-hydrolyzable esters (which may be hydrolyzedunder physiological conditions to release the compounds of the presentinvention in the form of free acids or alcohols). The compound of thepresent invention itself may be an ester as well.

The compound of the present invention can exist as a solvate (preferablya hydrate), wherein the compound of the present invention contains apolar solvent, in particular water, methanol or ethanol for example, asa structural element of the crystal lattice of the compound. The amountof the polar solvent, in particular water, may exist in a stoichiometricor non-stoichiometric ratio.

As can be appreciated by a person skilled in the art, not all nitrogencontaining heterocycles can form N-oxides since the nitrogen requires anavailable lone-pair electron for oxidation to the oxide; a personskilled in the art will recognize those nitrogen containing heterocycleswhich can form N-oxides. A person skilled in the art will also recognizethat tertiary amines can form N-oxides. Synthetic methods for thepreparation of N-oxides of heterocycles and tertiary amines are wellknown to a person skilled in the art, and they include the oxidation ofheterocycles and tertiary amines with peroxy acids such as peraceticacid and m-chloroperbenzoic acid (MCPBA), hydrogen peroxide, alkylhydroperoxides such as tert-butyl hydroperoxide, sodium perborate, anddioxiranes such as dimethyldioxirane. These methods for the preparationof N-oxides have been extensively described and reviewed in literatures,see e.g., T. L. Gilchrist, Comprehensive Organic Synthesis, vol. 7, pp748-750; A. R. Katritzky and A. J. Boulton, Eds., Academic Press; and G.W. H. Cheeseman and E. S. G. Werstiuk, Advances in HeterocyclicChemistry, vol. 22, pp 390-392, A. R. Katritzky and A. J. Boulton, Eds.,Academic Press.

The metabolite of the compound of the present invention, namely asubstance formed in vivo upon administration of the compound of thepresent invention, is also included within the scope of the presentinvention. Such a product may result e.g., from the oxidation,reduction, hydrolysis, amidation, de-amidation, esterification,enzymolysis, and the like, of the administered compound. Accordingly,the present invention encompasses the metabolite of the compound of thepresent invention, including a compound produced by a method comprisingcontacting the compound of the present invention with a mammal for aperiod of time sufficient to result in a metabolic product thereof.

Also within the scope of the present invention is a prodrug of thecompound of the invention, which is certain derivative of the compoundof the invention that may have little or no pharmacological activityitself, but can, when administered into or onto the body, be convertedinto the compound of the invention having the desired activity, forexample, by hydrolytic cleavage. In general, such prodrug will be afunctional derivative of the compound which is readily converted in vivointo the compound with desired therapeutic activity. Further informationon the use of the prodrug may be found in “Pro-drugs as Novel DeliverySystems”, Vol. 14, ACS Symposium Series (T. Higuchi and V. Stella). Theprodrug in accordance with the invention can, for example, be producedby replacing appropriate functionalities present in the compound of thepresent invention with certain moieties known to those skilled in theart as “pro-moieties” as described, for example, in “Design of Prodrugs”by H. Bundgaard (Elsevier, 1985).

The present invention further encompasses the compound of the presentinvention having a protecting group. During any of the processes forpreparation of the compound of the present invention, it may benecessary and/or desirable to protect sensitive or reactive groups onany of the molecules concerned, thereby resulting in the chemicallyprotected form of the compound of the present invention. This may beachieved by means of conventional protecting groups, e.g., thosedescribed in T. W. Greene & P. G. M. Wuts, Protective Groups in OrganicSynthesis, John Wiley & Sons, 1991, which is incorporated herein byreference. The protecting groups may be removed at a convenientsubsequent stage using methods known from the art.

The term “about” refers to a range within +10%, preferably within +5%,and more preferably within +2% of the specified value.

SPECIFIC EMBODIMENTS

Compound

In some embodiments, the present invention provides a compound or apharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein the compound has the structure of Formula (I):

wherein:

L is selected from the group consisting of C(═O), CRR′, NR, O, S, S═Oand S(═O)₂;

V¹ is selected from the group consisting of N,

and NR;

V² is selected from the group consisting of CR⁶ and C(═O);

represents either a single bond or a double bond, provided that when

is a single bond, V¹ is NR and V² is C(═O);

R and R′ are each independently selected from the group consisting of H,halogen, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated or partiallyunsaturated C₃₋₁₀ cyclic hydrocarbyl, saturated or partially unsaturated3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryland C₆₋₁₂ aralkyl, and at most 2 ring members in the cyclic hydrocarbyland heterocyclyl are C(═O);

R¹, R², R³ and R⁶ are each independently selected from the groupconsisting of H, halogen, —CN, —NH₂, —OH, —SH, —Se—R, —Si(R)₃, C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated or partially unsaturatedC₃₋₁₀ cyclic hydrocarbyl, saturated or partially unsaturated 3- to10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl,C₆₋₁₂ aralkyl, C₁₋₆ haloalkyl, —C(═O)R^(a), —OC(═O)R^(a), —C(═O)OR^(a),—OR^(a), —SR^(a), —S(═O)R^(a), —S(═O)₂R^(a), —S(═O)₂NR^(a)R^(b),—S(═O)(═NR)R^(a), —NR^(a)R^(b), —C(═O)NR^(a)R^(b), —C(═S)NR^(a)R^(b),—C(═NR)NR^(a)R^(b), —NR^(a)—C(═O)R^(b), —NR^(a)—C(═O)OR^(b),—NR^(a)—S(═O)₂—R^(b), —NR^(a)—C(═O)—NR^(a)R^(b), —C₁₋₆alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a), —C₁₋₆ alkylene-C(═O)R,—C₁₋₆ alkenylene-OR^(a), —O—C₁₋₆ alkylene-NR^(a)R^(b) and—P(═O)R^(a)R^(b);

R⁴ and R⁵ are each independently selected from the group consisting ofH, —C(═O)OR^(a), —NR^(a)R^(b), —NR^(a)—C(═O)R^(b), —NR^(a)—C(═O)OR^(b),—C₁₋₆ alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a), —C₁₋₆ alkylene-O—C₁₋₆alkylene-OR^(a), C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl,saturated or partially unsaturated C₃₋₁₀ cyclic hydrocarbyl, saturatedor partially unsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5-to 14-membered heteroaryl and C₆₋₁₂ aralkyl;

alternatively, R¹ and R⁴ together form —NH—(C₁₋₆ alkylene)-L-(C₁₋₆alkylene)-, preferably —NHCH₂CH₂—O—CH₂CH₂—;

the above alkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbyl,heterocyclyl, aryl, heteroaryl and aralkyl, at each occurrence, are eachoptionally substituted with one or more substituents independentlyselected from the group consisting of halogen, hydroxyl, oxo, amino,cyano, nitro, —Si(R)₃, C₁₋₆ alkyl, saturated or partially unsaturatedC₃₋₆ cyclic hydrocarbyl, saturated or partially unsaturated 3- to10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl,C₆₋₁₂ aralkyl, —C(═O)R^(a), —OC(═O)R^(a), —C(═O)OR^(a), —OR^(a),—SR^(a), —S(═O)R^(a), —S(═O)₂R^(a), —S(═O)₂NR^(a)R^(b), —NR^(a)R^(b),—C(═O)NR^(a)R^(b), —NR^(a)—C(═O)R^(b), —NR^(a)—C(═O)OR^(b),—NR^(a)—S(═O)₂—R^(b), —NR^(a)—C(═O)—NR^(a)R^(b), —C₁₋₆alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a), —C₁₋₆ alkenylene-OR^(a) and—O—C₁₋₆ alkylene-NR^(a)R^(b), the alkyl, cyclic hydrocarbyl,heterocyclyl, aryl, heteroaryl and aralkyl are further optionallysubstituted with one or more substituents independently selected fromthe group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro,—NR^(a)R^(b), C₁₋₆ alkyl, —O—C₁₋₆ alkyl, saturated or partiallyunsaturated C₃₋₆ cyclic hydrocarbyl, saturated or partially unsaturated3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryland C₆₋₁₂ aralkyl; and R^(a) and R^(b), at each occurrence, are eachindependently selected from the group consisting of H, —OH, C₁₋₆ alkyl,C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated or partially unsaturated C₃₋₁₀cyclic hydrocarbyl, saturated or partially unsaturated 3- to 10-memberedheterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl and C₆₋₁₂aralkyl; alternatively, R^(a) and R^(b) together with the atom to whichthey are attached form a 3- to 12-membered heterocycle or heteroaromaticring, the above groups are further optionally substituted with one ormore substituents independently selected from the group consisting ofhalogen, hydroxyl, oxo, amino, cyano, nitro, C₁₋₆ alkyl, —O—C₁₋₆ alkyl,saturated or partially unsaturated C₃₋₆ cyclic hydrocarbyl, saturated orpartially unsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to14-membered heteroaryl and C₆₋₁₂ aralkyl.

In preferred embodiments, L is selected from the group consisting ofCH₂, O, S and NH.

In preferred embodiments, V¹ is selected from the group consisting of N,

and NCH₃.

In preferred embodiments, V² is selected from the group consisting ofCH, C—NHCH₃, C—OCH₃, C—F and C(═O).

In preferred embodiments, the present invention provides the compound ora pharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein R^(a) and R^(b), at each occurrence, are eachindependently selected from the group consisting of H, —OH, methyl,ethyl, n-propyl, isopropyl, cyclopropyl, phenyl, benzyl, methoxy andethoxy; alternatively, R^(a) and R^(b) together with the atom to whichthey are attached form a 5- to 8-membered heterocycle or heteroaromaticring.

In preferred embodiments, the present invention provides the compound ora pharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein R¹, R², R³ and R⁶ are each independently selected fromthe group consisting of H, F, Cl, Br, I, —CN, —NH₂, —OH, —SH, —Se—CH₃,—Si(CH₃)₃, —CH₂NH₂, —CH₂NHCH₃, —CH₂N(CH₃)₂, methyl, ethyl, n-propyl,isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, vinyl, propenyl, allyl, ethynyl, propynyl,trifluoromethyl, acetyl, —C(═O)OH, —C(═O)NH₂, —C(═S)NH₂, —C(═NH)NH₂,—NHCH₃, —NHCH₂CH₃, —NHCH₂CF₃, —N(CH₃)₂, —N(CH₃)(C₂H₅), —N(C₂H₅)₂,—NHCH₂CH₂OH, —NH—C(═O)CH₃, —NH—C(═O)CH═CH₂, methoxy, ethoxy, propoxy,phenyl, —NH—C(═O)—NH₂, —NH—C(═O)OCH₃, —SCH₃, —SCH₂CH₃, —SC(CH₃)₃, —SBn,—S(═O)CH₃, —S(═O)Bn, —S(═O)₂CH₃, —S(═O)₂Bn, —S(═O)₂NH₂, —S(═O)₂NHCH₃,—S(═O)₂N(CH₃)₂, —S(═O)(═NH)CH₃, —P(═O)(CH₃)₂, —P(═O)(C₂H₅)₂,

In preferred embodiments, the present invention provides the compound ora pharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein R⁴ and R⁵ are each independently selected from thegroup consisting of H, —C(═O)OC(CH₃)₃, —NH₂, —NHCH₃, —NHPh, —NHC(═O)CH₃,—NHBoc, methyl, ethyl, —CH₂CF₃, n-propyl, isopropyl, n-butyl, isobutyl,tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, phenyl,benzyl,

In preferred embodiments, the present invention provides the compound ora pharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein the compound has the structure of any of the followingformulae:

preferably has the structure of any of the following formulae:

The compound obtained by any combination of the various embodiments isencompassed by the invention.

In preferred embodiments, the present invention provides the compound ora pharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, wherein the compound has the following structure:

Pharmaceutical Composition and Therapeutic Method

In some embodiments, the present invention provides a pharmaceuticalcomposition comprising a prophylactically or therapeutically effectiveamount of the compound of the present invention or a pharmaceuticallyacceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide,isotopically labeled compound, metabolite or prodrug thereof and one ormore pharmaceutically acceptable carriers, and the pharmaceuticalcomposition is preferably in the form of a solid, semi-solid, liquid, orgas preparation. In some embodiments, the pharmaceutical composition canfurther comprise one or more additional therapeutic agents.

In some embodiments, the present invention provides use of the compoundof the present invention or a pharmaceutically acceptable salt, ester,stereoisomer, polymorph, solvate, N-oxide, isotopically labeledcompound, metabolite or prodrug thereof or the pharmaceuticalcomposition of the present invention in the manufacture of a medicamentfor the treatment of a disease mediated by the P2X3 and/or P2X2/3receptor antagonist.

In some embodiments, the present invention provides the compound of thepresent invention or a pharmaceutically acceptable salt, ester,stereoisomer, polymorph, solvate, N-oxide, isotopically labeledcompound, metabolite or prodrug thereof or the pharmaceuticalcomposition of the present invention for use in the treatment of adisease mediated by the P2X3 and/or P2X2/3 receptor antagonist.

In some embodiments, the present invention provides a method for theprophylaxis or the treatment of a disease mediated by the P2X3 and/orP2X2/3 receptor antagonist, wherein the method comprises administeringto a subject in need thereof an effective amount of the compound of thepresent invention or a pharmaceutically acceptable salt, ester,stereoisomer, polymorph, solvate, N-oxide, isotopically labeledcompound, metabolite or prodrug thereof or the pharmaceuticalcomposition of the present invention.

In some embodiments, the disease mediated by the P2X3 and/or P2X2/3receptor antagonist is selected from the group consisting of a urinarytract disease selected from reduced bladder capacity, frequentmicturition, urge incontinence, stress incontinence, bladderhyperreactivity, benign prostatic hypertrophy, prostatitis, detrusorhyperreflexia, nocturia, urinary urgency, pelvic hypersensitivity,urethritis, pelvic pain syndrome, prostatodynia, cystitis, andidiopathic bladder hypersensitivity; pain disease selected frominflammatory pain, surgical pain, visceral pain, dental pain,premenstrual pain, central pain, pain due to burns, migraine and clusterheadaches, nerve injury, neuritis, neuralgia, poisoning, ischemicinjury, interstitial cystitis, cancer pain, viral, parasitic orbacterial infection, post-traumatic injury and pain associated withirritable bowel syndrome; cardiovascular system disease, preferablyhypertension; respiratory disease selected from chronic obstructivepulmonary disease, asthma and bronchospasm; gastrointestinal diseaseselected from irritable bowel syndrome (preferably diarrhea-dominantirritable bowel syndrome), inflammatory bowel disease, biliary colic,renal colic, and pain associated with gastrointestinal distension.

The term “pharmaceutically acceptable carrier” in the present inventionrefers to a diluent, auxiliary material, excipient, or vehicle withwhich a therapeutic is administered, and it is, within the scope ofsound medical judgment, suitable for contact with the tissues of humanbeings and animals without excessive toxicity, irritation, allergicresponse, or other problem or complication, commensurate with areasonable benefit/risk ratio.

The pharmaceutically acceptable carrier which can be employed in thepharmaceutical composition of the present invention includes, but is notlimited to sterile liquids, such as water and oils, including those ofpetroleum, animal, vegetable or synthetic origin, such as peanut oil,soybean oil, mineral oil, sesame oil and the like. Water is an exemplarycarrier when the pharmaceutical composition is administeredintravenously. Physiological salines as well as aqueous dextrose andglycerol solutions can also be employed as liquid carriers, particularlyfor injectable solutions. Suitable pharmaceutical excipients includestarch, glucose, lactose, sucrose, gelatin, maltose, chalk, silica gel,sodium stearate, glycerol monostearate, talc, sodium chloride, driedskim milk, glycerol, propylene glycol, water, ethanol and the like. Thepharmaceutical composition, if desired, can also contain minor amountsof wetting or emulsifying agents, or pH buffering agents. Oralformulations can include standard carriers such as pharmaceutical gradesof mannitol, lactose, starch, magnesium stearate, sodium saccharine,cellulose, magnesium carbonate, etc. Examples of suitable pharmaceuticalcarriers are described in e.g. Remington's Pharmaceutical Sciences(1990).

The pharmaceutical composition of the present invention can actsystemically and/or topically. To this end, it can be administeredthrough a suitable route, such as through injection, (intravenous,intraarterial, subcutaneous, intraperitoneal, intramuscular injection,including dripping), or transdermal administration, or administered viaoral, buccal, nasal, transmucosal, topical, as an ophthalmicformulation, or via inhalation.

For these routes of administration, the pharmaceutical composition ofthe present invention can be administered in a suitable dosage form.

Such dosage forms include, but are not limited to tablets, capsules,lozenges, hard candies, powders, sprays, creams, salves, suppositories,gels, pastes, lotions, ointments, aqueous suspensions, injectablesolutions, elixirs, and syrups.

As used herein, the term “effective amount” refers to the amount of acompound being administered which will relieve to some extent one ormore of the symptoms of the disorder being treated.

Dosage regimens may be adjusted to provide the optimum desired response.For example, a single bolus may be administered, several divided dosesmay be administered over time, or the dose may be proportionally reducedor increased as indicated by the exigencies of the therapeuticsituation. It is to be noted that dosage values may vary with the typeand severity of the condition to be alleviated, and may include singleor multiple doses. It is to be further understood that for anyparticular subject, specific dosage regimens should be adjusted overtime according to the individual need and the professional judgment ofthe person administering or supervising the administration of thecomposition.

The amount of the compound of the present invention administered will bedependent on the subject being treated, the severity of the disorder orcondition, the rate of administration, the disposition of the compoundand the discretion of the prescribing physician. Generally, an effectivedosage is in the range of about 0.0001 to about 50 mg per kg body weightper day, for example about 0.01 to about 10 mg/kg/day, in single ordivided doses. For a 70 kg human, this would amount to about 0.007 mg toabout 3500 mg/day, for example about 0.7 mg to about 700 mg/day. In someinstances, dosage levels below the lower limit of the aforesaid rangemay be more than adequate, while in other cases, still larger doses maybe employed without causing any harmful side effect, provided that suchlarger doses are first divided into several small doses foradministration throughout the day.

The content or dosage of the compound of the present invention in thepharmaceutical composition is about 0.01 mg to about 1000 mg, suitably0.1-500 mg, preferably 0.5-300 mg, more preferably 1-150 mg,particularly preferably 1-50 mg, e.g., 1.5 mg, 2 mg, 4 mg, 10 mg, 25 mg,etc.

Unless otherwise indicated, the term “treating” or “treatment”, as usedherein, means reversing, alleviating, inhibiting the progress of, orpreventing the disorder or condition to which such term applies, or oneor more symptoms of such disorder or condition.

As used herein, the term “subject” includes a human or non-human animal.An exemplary human subject includes a human subject having a disease(such as one described herein) (referred to as a patient), or a normalsubject. The term “non-human animal” as used herein includes allvertebrates, such as non-mammals (e.g. birds, amphibians, reptiles) andmammals, such as non-human primates, livestock and/or domesticatedanimals (such as sheep, dog, cat, cow, pig and the like).

In some embodiments, the pharmaceutical composition of the presentinvention can further comprise one or more additional therapeutic agentsor prophylactic agents.

EXAMPLES

The present invention is further described with reference to thefollowing examples, which are not provided to limit the scope of thepresent invention.

The structure of the compound was confirmed by nuclear magneticresonance spectrum (¹H NMR) or mass spectrum (MS).

Chemical shifts (S) are expressed in parts per million (ppm). ¹H NMR wasrecorded on a Bruker 400 or Varian 300 spectrometer, the test solventwas deuterated methanol (CD₃OD), deuterated chloroform (CDCl₃) orhexadeuterated dimethyl sulfoxide (DMSO-d₆), and the internal standardwas tetramethylsilane (TMS).

The LC-MS assay was conducted on Agilent LC-MS-1110 liquidchromatography-mass spectrometer, Agilent LC-MS-6110 liquidchromatography-mass spectrometer, Agilent LC-MS-6120 liquidchromatography-mass spectrometer (Manufacturer: Agilent) or ShimadzuLC-MS-2020.

Preparative high-performance liquid chromatography was conducted on MSinduced AutoPurification system (Waters), Gilson GX-281 (Gilson), orsemi-preparative liquid chromatograph (Tong Heng Innovation TechnologyCo., Ltd., LC3000 (Ddlsogel, C18, 30 mm×250 mm 10 μm).

Thin layer chromatography (TLC) was performed with Huanghai HSGF 254(5×20 cm) silica gel plates, and preparative thin layer chromatographywas performed with GF 254 (0.4˜0.5 nm) silica gel plates produced inYantai.

The reaction was monitored by thin layer chromatography (TLC) or LC-MS,the developing solvent system included dichloromethane and methanolsystem, n-hexane and ethyl acetate system, as well as petroleum etherand ethyl acetate system, and was adjusted (by adjusting the volumeratio of the solvents, or by adding triethylamine, etc.) according tothe polarity of the compound to be separated.

The microwave reaction was conducted by CEM Discovery Sp (400 W, RT˜300°C.) microwave reactor.

Silica gel (200˜300 mesh) produced by Yucheng Chemical Co., Ltd wasnormally employed as a stationary phase in column chromatography. Theeluent system included dichloromethane and methanol system, as well asn-hexane and ethyl acetate system, and was adjusted (by adjusting thevolume ratio of the solvents, or by adding triethylamine, etc.)according to the polarity of the compound to be separated.

In the following examples, unless otherwise specified, the reactiontemperature was room temperature (20° C.˜30° C.).

The reagents employed in the Examples were purchased from companies suchas Aldrich Chemical Company, Shanghai Bide Pharmatech Co. Ltd., BeijingGreenchem Co. Ltd., Shanghai Shaoyuan Co. Ltd. or Ables Technology Co.Ltd. etc.

The abbreviations as used in the present invention have the followingmeanings:

Abbreviation Meaning ACN acetonitrile BBr₃ boron tribromide BnBr benzylbromide Br₂ bromine CBr₄ carbon tetrabromide DCM dichloromethaneDIEA/DIPEA N,N-diisopropylethylamine DMA-DMF N,N-dimethylformamidedimethyl acetal DMAP 4-dimethylaminopyridine DMF N,N-dimethylformamideEA ethyl acetate H₂ hydrogen HBr hydrobromic acid HCl hydrochloric acidH₂O water K₂CO₃ potassium carbonate LDA lithium diisopropylamide m-CPBAmeta-chloroperbenzoic acid MeCN acetonitrile MeOH methanol MeONa sodiummethoxide Na₂CO₃ sodium carbonate NaH sodium hydride NaOH sodiumhydroxide NaSMe sodium thiomethoxide n-BuLi n-butyl lithium NCSN-chlorosuccinimide NMP N-methylpyrrolidone Oxone potassiumperoxymonosulfate Pd/C palladium/carbon Pd₂(dba)₃tris(dibenzylideneacetone)dipalladium Pd(dppf)Cl₂[1,1’-bis(diphenylphosphino)ferrocene]dichloropalladium Pd(PPh₃)₄tetrakis(triphenylphosphine)palladium Pd(PPh₃)₂Cl₂bis(triphenylphosphine)dichloropalladium PE petroleum ether PMBClp-methoxybenzyl chloride TBAF tetrabutylammonium fluoride TEAtriethylamine TFA trifluoroacetic acid TFAA trifluoroacetic anhydrideTHF tetrahydrofuran Xantphos4,5-bis(diphenylphosphino)-9,9-dimethylxanthene x-phos 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl

Example 1: preparation of5-((2-bromo-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C2,compound 2)

Step 1:

Compound C2-1 (100 g, 0.54 mol) was dissolved in 1,4-dioxane (700 mL),the starting material SM2 (136 g, 0.81 mol), K₂CO₃ (149 g, 1.08 mol) andPd(PPh₃)₄ (6.2 g, 5.4 mmol) were sequentially added, followed byaddition of purified water (35 mL), and purge with nitrogen wasperformed for 3 times. Under the protection of nitrogen, the reactionwas performed at 100° C. for 18 hours. LC-MS indicated the reaction ofthe starting materials was substantially complete. The reaction solutionwas cooled to room temperature, filtered, and the filter cake was washedwith 1,4-dioxane (200 mL). The filtrate was concentrated under reducedpressure to remove 1,4-dioxane, followed by addition of purified water(200 mL), and extraction with ethyl acetate (400 mL×3). The organicphases were combined, dried over anhydrous sodium sulfate (100 g) for 30min, filtered, and concentrated under reduced pressure to afford a crudeproduct. The crude product was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=20:1˜10:1), to afford compoundC2-2 (79 g, yellow oil, yield: 99.75%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.37 (d, J=5.6 Hz, 1H), 8.22 (s, 1H), 7.04(d, J=5.6 Hz, 1H), 5.18 (s, 1H), 5.09 (s, 1H), 3.85 (s, 3H), 2.05 (s,3H); MS m/z (ESI): 150.0 [M+H]⁺.

Step 2:

Compound C2-2 (79 g, 0.53 mol) was dissolved in anhydrous methanol (700mL), 10% palladium/carbon (16 g) was added, and the reaction wasperformed under hydrogen (0.4 MPa) at room temperature for 18 hours.LC-MS indicated a small amount of the starting material remained.palladium/carbon (4 g) was supplemented, and the reaction was continuedunder hydrogen (0.4 MPa) at room temperature for 18 hours. LC-MSindicated the reaction of the starting materials was complete. Thereaction solution was filtered, the filter cake was washed with methanol(100 mL), and the filtrate was concentrated under reduced pressure togive a crude product, compound C2-3 (80 g, orange oily liquid, yield:99.96%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.31 (d, J=5.6 Hz, 1H), 8.28 (s, 1H), 6.98(d, J=5.6 Hz, 1H), 3.86 (s, 3H), 3.21-3.09 (m, 1H), 1.21 (d, J=7.2 Hz,6H); MS m/z (ESI): 152.1 [M+H]⁺.

Step 3:

Compound N,N-dimethylethanolamine (46.3 g, 0.52 mol) was dissolved inn-hexane (400 mL). Under the protection of nitrogen, the reaction wascooled to −15° C.˜−20° C., 2.4 M/L n-butyl lithium (434 mL, 1.04 mol)was slowly dropwise added. After the dropwise addition was complete, thereaction was kept at the temperature for 30 minutes, and then a solutionof compound C2-3 (40 g, 0.26 mol) in toluene (200 mL) was slowlydropwise added at −15° C.˜−20° C. After the dropwise addition wascomplete, the reaction was kept at the temperature for 30 minutes. Thereaction solution was cooled to −70° C., a solution of carbontetrabromide (172.4 g, 0.52 mol) in toluene (500 mL) was slowly dropwiseadded, and the temperature was controlled at −70° C.˜−75° C. After thedropwise addition was complete, the reaction was kept at the temperaturefor 1 hour. LC-MS indicated the reaction of the starting materials wascomplete. The reaction was quenched by adding water (500 mL), andextracted with ethyl acetate (500 mL×3). The organic phases werecombined, washed once with saturated brine (500 mL), dried overanhydrous sodium sulfate (400 g) for half an hour, filtered andconcentrated. The crude product was isolated by column chromatography onsilica gel (petroleum ether:ethyl acetate=200:1˜50:1) to afford compoundC2-4 (25 g, light yellow oily liquid, yield: 41.81%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.06 (s, 1H), 7.20 (s, 1H), 3.89 (s, 3H),3.13-3.05 (m, 1H), 1.18 (d, J=6.8 Hz, 6H); MS m/z (ESI): 229.9 [M+H]⁺.

Step 4:

Compound C2-4 (25 g, 0.11 mol) was dissolved in dichloromethane (300mL). Under the protection of nitrogen, the reaction was cooled to 0°C.˜5° C., and a solution of boron tribromide (140.3 g, 0.55 mol) wasslowly added. After completion of the addition, the reaction solutionwas warmed to reflux, and the reaction was performed for 18 hours. LC-MSindicated the reaction of the starting materials was complete.

The reaction solution was cooled to room temperature, and slowlydropwise added to 500 g ice. After the dropwise addition was complete, asaturated solution of sodium bicarbonate was dropwise added to adjust pHto 7˜8. The reaction was filtered, the filter cake was washed thricewith ethyl acetate (400 mL), the filtrate was separated, and the aqueousphase was extracted with ethyl acetate (400 mL×3) again. All the organicphases were combined, dried over anhydrous sodium sulfate (500 g) forhalf an hour, filtered, and the filtrate was concentrated under reducedpressure to afford compound C2-5 (20 g, light yellow solid, yield:84.17%).

¹H NMR (400 MHz, DMSO-d₆) δ 11.11 (s, 1H), 7.99 (s, 1H), 6.90 (s, 1H),3.10-3.02 (m, 1H), 1.18 (d, J=6.8 Hz, 6H); MS m/z (ESI): 215.9 [M+H]⁺.

Step 5:

Compound C2-5 (10 g, 0.047 mol) was dissolved in DMF (50 mL). Under theprotection of nitrogen, potassium carbonate (12.8 g, 0.093 mol) andbromoacetonitrile (8.4 g, 0.07 mol) were sequentially added, and thereaction was stirred at room temperature for 2 hours. LC-MS indicatedthe reaction of the starting materials was complete. The reaction wasquenched by adding water (50 mL), and extracted with ethyl acetate (50mL×4). The combined organic phases were washed with saturated brine (50mL×3), added with anhydrous sodium sulfate, dried for half an hour, andfiltered. The filtrate was concentrated under reduced pressure, and thecrude product was isolated by column chromatography on silica gel(petroleum ether:ethyl acetate=20:1˜5:1) to afford compound C2-6 (4 g,light yellow solid, yield: 33.38%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.18 (s, 1H), 7.40 (s, 1H), 5.37 (s, 2H),3.14-3.06 (m, 1H), 1.21 (d, J=6.8 Hz, 6H); MS m/z (ESI): 254.8 [M+H]⁺.

Step 6:

Compound C2-6 (4 g, 0.016 mol) was dissolved in DMF (50 mL). Under theprotection of nitrogen, tert-butoxy bis(dimethylamino)methane (8.2 g,0.048 mol) was added, the reaction was heated to 100° C., and stirredfor 2 hours. LC-MS indicated the reaction of the starting materials wascomplete. The reaction solution was cooled to room temperature, quenchedby adding water (50 mL), and then extracted with ethyl acetate (50mL×3). The organic phase was then washed with saturated brine (50 mL×3),added with anhydrous sodium sulfate, dried for half an hour, andfiltered. The filtrate was concentrated under reduced pressure, and thecrude product was isolated by column chromatography on silica gel(petroleum ether:ethyl acetate=10:1˜5:1) to afford compound C2-7 (3.8 g,light yellow solid, yield: 66.90%). MS m/z (ESI): 309.7 [M−45+H]*.

Step 7:

Compound C2-7 (3.54 g, 0.01 mol) was dissolved in DMF (25 mL). Under theprotection of nitrogen, aniline hydrobromide (2.08 g, 0.012 mol) wasadded, the reaction was heated to 100° C., and stirred for 2 hours.LC-MS indicated the reaction of the starting materials was complete. Thereaction solution was cooled to room temperature, quenched by addingwater (25 mL), and extracted with ethyl acetate (20 mL×3). The organicphase was then washed with saturated brine (20 mL×3), added withanhydrous sodium sulfate, dried for half an hour, and filtered. Thefiltrate was concentrated under reduced pressure, and the crude productwas isolated by column chromatography on silica gel (petroleumether:ethyl acetate=20:1˜5:1) to afford compound C2-8 (3.1 g, lightyellow solid, yield: 86.59%).

¹H NMR (400 MHz, DMSO-d₆) δ 9.36 (d, J=12.8 Hz, 1H), 8.28 (s, 1H), 7.95(d, J=12.8 Hz, 1H), 7.32-7.24 (m, 4H), 7.20 (s, 1H), 6.99 (t, J=7.2 Hz,1H), 3.31-3.26 (m, 1H), 1.28 (d, J=6.8 Hz, 6H); MS m/z (ESI): 357.7[M+H]⁺.

Step 8:

Guanidine hydrochloride (2.4 g, 25.2 mmol) was added to anhydrousethanol (50 mL). Under the protection of nitrogen, sodium methoxide (2.4g, 25.2 mmol) was added, the reaction was stirred at room temperaturefor half an hour, followed by addition of compound C2-8 (3 g, 8.4 mmol).After completion of the addition, the reaction solution was heated toreflux, and the reaction was performed for 18 hours. LC-MS indicated thereaction of the starting materials was complete. The reaction solutionwas cooled to room temperature, filtered, the filtrate was concentratedunder reduced pressure, and the crude product was isolated by columnchromatography on silica gel (DCM:MeOH=50:1˜20:1) to afford compound C2(900 mg, light yellow solid, yield: 33.17%, compound 2).

¹H NMR (400 MHz, DMSO-d₆) δ 8.19 (s, 1H), 7.62 (s, 1H), 6.56 (s, 1H),6.47 (s, 2H), 6.06 (s, 2H), 3.32-3.27 (m, 1H), 1.28 (d, J=6.8 Hz, 6H);MS m/z (ESI): 323.7 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 1.

Starting material or regent different Compound Compound from that in No.Structure Name Example 1 Characterization Data C3 (compound 3)

5-((2-chloro-5- isopropylpyridin- 4- yl)oxy)pyrimidine- 2,4-diamineCarbon tetrabromide in step 3 was replaced with hexachloroethane, andaniline hydrobromide in Step 7 was replaced with aniline hydrochloride.¹H NMR (400 MHz, DMSO-d₆) δ 8.21 (s, 1H), 7.64 (s, 1H), 6.53 (s, 2H),6.48 (s, 1H), 6.11 (s, 2H), 3.32-3.29 (m, 1H), 1.29 (d, J = 6.8 Hz, 6H);MS m/z (ESI): 279.8 [M + H]⁺. C12 (compound 12)

5-((2-iodo-5- isopropylpyridin- 4- yl)oxy)pyrimidine- 2,4-diamine Carbontetrabromide in Step 3 was replaced with iodine, and anilinehydrobromide in Step 7 was replaced with aniline hydroiodide. ¹H NMR(400 MHz, DMSO-d₆) δ 8.15 (s, 1H), 7.62 (s, 1H), 6.77 (s, 2H), 6.54 (s,1H), 6.13 (s, 2H), 3.32- 3.29 (m, 1H), 1.27 (d, J = 6.8 Hz, 6H); MS m/z(ESI): 371.7 [M + H]⁺. C67 (compound 67)

5-((5-isopropyl- 2- (trifluoromethyl) pyridin-4- yl)oxy)pyrimidine-2,4-diamine C2-5 in Step 5 was replaced with C67- 4*. ¹H NMR (400 MHz,CD₃OD) δ 8.63 (s, 1H), 7.82 (s, 1H), 7.30 (s, 1H), 3.53-3.49 (m, 1H),1.43-1.41 (d, J = 7.2 Hz, 6H). MS m/z (ESI): 313.9 [M + H]⁺. C215-1

5-((2-bromo-5- (prop-1-en-2- yl)pyridin-4- yl)oxy)pyrimidine-2,4-diamine Step 2 in Example 1 was omitted. MS m/z (ESI): 321.7 [M +H]⁺. *preparation of intermediate C67-4:

Step 1:

Compound C67-1 (10 g, 0.061 mol) was dissolved in acetic acid (100 mL),bromine (11.7 g, 0.073 mol) was dissolved in an aqueous solution ofpotassium hydroxide (10.3 g, 0.183 mol, 20 mL), which was then slowlyadded dropwise to the reaction solution under ice bath cooling. Thereaction solution was stirred overnight at room temperature. LC-MSindicated the reaction of the starting materials was substantiallycomplete. The reaction was rotary evaporated to remove acetic acid, thenadjusted to pH 7 with sodium bicarbonate, and extracted with ethylacetate (200 mL×3). The organic phases were combined, washed once withsaturated brine (100 mL), then dried over anhydrous sodium sulfate (50g) for half an hour, filtered, and concentrated under reduced pressureto afford a crude product. The crude product was isolated by columnchromatography on silica gel (petroleum ether:ethyl acetate=5:1) toafford compound C67-2 (6 g, white solid, yield 40.5%).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (s, 1H), 6.84 (s, 1H). MS m/z (ESI):241.7 [M+H]⁺.

Step 2:

Compound C67-2 (6 g, 0.025 mol) was dissolved in 1,4-dioxane (100 mL),and isopropenyl pinacol borate (6.27 g, 0.037 mol), potassium carbonate(6.87 g, 0.050 mol), Pd(PPh₃)₄ (0.6 g) and water (10 mL) were added.Purge with nitrogen was performed for 3 times, and the reaction solutionwas stirred at 100° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was cooled toroom temperature, quenched by adding water (100 mL), and extracted withethyl acetate (100 mL×3). The organic phases were combined, then washedwith saturated brine (100 mL), added with anhydrous sodium sulfate (50g), dried for half an hour, and filtered. The filtrate was concentratedunder reduced pressure to afford a crude product, which was isolated bycolumn chromatography on silica gel (petroleum ether:ethylacetate=10:1˜5:1) to afford compound C67-3 (1 g, yellow oily liquid,yield 20%). MS m/z (ESI): 203.9 [M+H]⁺.

Step 3:

Compound C67-3 (1 g, 0.0049 mol) was dissolved in methanol (10 mL), wetpalladium/carbon (0.3 g) was added, and the reaction solution wasstirred at room temperature for 16 hours. LC-MS indicated the reactionof the starting materials was substantially complete. The reaction wasfiltered, concentrated under reduced pressure to afford a crude product,C67-4, which was directly used in the next step (1 g, light yellow oilyliquid, yield 100%). MS m/z (ESI): 205.9 [M+H]⁺.

Example 2: preparation of5-((5-isopropyl-2-methoxypyridin-4-yl)oxy)pyrimidine-2,4-diamine(Compound C4, Compound 4)

Step 1:

Compound C2-5 (440 mg, 2.03 mmol) and methanol (6 mL) was added to a 50mL flask, sodium methoxide (220 mg, 4.06 mmol) was added, followed byslow dropwise addition of benzyl bromide (694 mg, 4.06 mmol), and thereaction was performed at room temperature overnight. Thin layerchromatography (ethyl acetate) and LC-MS analysis indicated the reactionwas complete. The reaction solution was directly used in the next step.MS m/z (ESI): 305.8/307.8 [M+H]⁺.

Step 2:

Sodium methoxide (220 mg, 4.06 mmol) was added to the reaction solutionobtained in the previous step, and the reaction was performed undermicrowave radiation at 100° C. for 1 hour. Thin layer chromatography(ethyl acetate) and LC-MS analysis indicated the reaction was complete.The reaction solution was cooled to room temperature, concentrated underreduced pressure to give a crude product, which was isolated bypreparative chromatography (ethyl acetate˜dichloromethane:methanol=10:1)to afford compound C4-3 (470 mg, colorless oil, yield: 75.66%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.46 (s, 1H), 7.38-7.28 (m, 3H), 7.18-7.16(m, 2H), 5.63 (s, 1H), 5.06 (s, 2H), 3.76 (s, 3H), 2.98-2.87 (m, 1H),1.07 (d, J=6.8 Hz, 6H); MS m/z (ESI): 257.9 [M+H]⁺.

Step 3:

In a 50 mL flask, compound C4-3 (470 mg, 1.83 mmol) was dissolved inmethanol (15 mL), 10% Pd/C (200 mg) was added, purge with hydrogen wasperformed 3 times, and the reaction was performed under hydrogenovernight. LC-MS analysis indicated the reaction was complete. Thereaction solution was filtered, the filter cake was rinsed with methanol(15 mL), and the filtrate was concentrated under reduced pressure toafford compound C4-4 (270 mg, white solid, yield: 88.35%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.50 (bs, 1H), 7.76 (s, 1H), 6.11 (s, 1H),3.74 (s, 3H), 3.04-2.97 (m, 1H), 1.17 (d, J=6.4 Hz, 6H); MS m/z (ESI):168.0 [M+H]⁺.

Step 4:

In a 50 mL flask, compound C4-4 (260 mg, 1.56 mmol) was dissolved inN,N-dimethylformamide (5 mL), potassium carbonate (430 mg, 3.11 mmol)was added, followed by slow dropwise addition of bromoacetonitrile (374mg, 3.11 mmol), and the reaction was performed at room temperature for 1hour. LC-MS analysis indicated the reaction was complete. The reactionwas quenched by water (10 mL), and then extracted with ethyl acetate (20mL) for 4 times. The organic phases were combined, washed with saturatedbrine (10 mL) 1 times, then dried over sodium sulfate, filtered, andconcentrated to give a crude product, which was purified by preparativechromatography (petroleum ether:ethyl acetate=3:1˜1:1) to affordcompound C4-5 (220 mg, colorless oil, yield: 68.45%). MS m/z (ESI):207.0 [M+H]⁺.

Step 5:

In a 50 mL flask, compound C4-5 (170 mg, 0.83 mmol) was dissolved inN,N-dimethylformamide (5 mL), tert-butoxy bis(dimethylamino)methane (435mg, 2.5 mmol) was added, the reaction solution was heated to 100° C.,and the reaction was performed for 18 hours. LC-MS analysis indicatedthe reaction was complete. The reaction was quenched by water (10 mL),and then extracted with ethyl acetate (20 mL) for 4 times. The organicphases were combined, washed with saturated brine (10 mL×2), then driedover sodium sulfate, filtered, and concentrated under reduced pressure.The crude product was purified by preparative chromatography (petroleumether:ethyl acetate=3:1˜1:1) to afford compound C4-6 (100 mg, colorlessoil, yield: 39.37%). MS m/z (ESI): 307.0 [M+H]⁺.

Step 6:

In a 50 mL flask, compound C4-6 (80 mg, 0.26 mmol) was dissolved inN,N-dimethylformamide (2 mL), aniline hydrochloride (40 mg, 0.29 mmol)was added, and the reaction was heated to 100° C., and stirred for 2hours. LC-MS indicated the reaction of the starting materials wascomplete. The reaction was added with 25 mL water, and extracted withethyl acetate 20 mL×3. Then, the organic phase was washed with saturatedbrine (20 mL×3), added with anhydrous sodium sulfate, dried for half anhour, and filtered. The filtrate was concentrated under reducedpressure, and the crude product was purified by preparativechromatography (petroleum ether:ethyl acetate=2:1) to afford compoundC4-7 (40 mg, colorless oil, yield: 49.79%). MS m/z (ESI): 310.0 [M+H]⁺.

Step 7:

In a 50 mL flask, compound C4-7 (40 mg, 0.13 mmol) was dissolved inanhydrous ethanol (2 mL), guanidine carbonate (29 mg, 0.29 mmol) wasadded, and the reaction was performed under microwave radiation at 100°C. for 2 hours. LC-MS indicated the reaction of the starting materialswas complete. The reaction was cooled to room temperature, andconcentrated under reduced pressure. The crude product was purified bypreparative chromatography (dichloromethane:methanol=10:1) to affordcompound C4 (20 mg, white solid, yield: 55.94%, compound 4).

¹H NMR (400 MHz, DMSO-d₆) δ 7.95 (s, 1H), 7.60 (s, 1H), 6.65 (s, 2H),6.26 (s, 2H), 5.85 (s, 1H), 3.76 (s, 3H), 3.28-3.22 (m, 1H), 1.26 (d,J=6.8 Hz, 6H); MS m/z (ESI): 275.9 [M+H]⁺.

Example 3: preparation of4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpyridin-2-ol (Compound C7,Compound 7)

Step 1:

In a 50 mL flask, compound C4-6 (25 mg, 0.08 mmol) was dissolved inN,N-dimethylformamide (2 mL), aniline hydrochloride (12 mg, 0.09 mmol)was added, the reaction was heated to 100° C., and stirred for 2 hours.LC-MS indicated the reaction of the starting materials was complete. Thereaction was added with water (25 mL), and extracted with ethyl acetate(20 mL×3). The organic phase was then washed with saturated brine (20mL×3), added with anhydrous sodium sulfate, dried for half an hour, andfiltered. The filtrate was concentrated under reduced pressure, and thecrude product was purified by preparative thin layer chromatography(petroleum ether:ethyl acetate=2:1) to afford compound C7-2 (15 mg,colorless oil, yield: 63.56%). MS m/z (ESI): 295.9 [M+H]⁺.

Step 2:

In a 50 mL flask, compound C7-2 (40 mg, 0.13 mmol) was dissolved inanhydrous ethanol (2 mL), guanidine carbonate (29 mg, 0.29 mmol) wasadded, and the reaction was performed under microwave radiation at 100°C. for 2 hours. LC-MS indicated the reaction of the starting materialswas complete. The reaction was cooled to room temperature, andconcentrated under reduced pressure. The crude product was purified bypreparative chromatography (dichloromethane:methanol=10:1) to affordcompound C7 (2 mg, white solid, yield: 55.89%, compound 7).

¹H NMR (400 MHz, CD₃OD) δ 7.64 (s, 1H), 7.25 (s, 1H), 5.67 (s, 1H),3.23-3.18 (m, 1H), 1.30 (d, J=6.8 Hz, 6H); MS m/z (ESI): 261.9 [M+H]⁺.

Example 4: preparation of5-((3-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C8, compound 8)

Step 1:

Compound C2 (20 mg, 0.06 mmol) was dissolved in anhydrous methanol (5mL), followed by addition of 10% palladium/carbon (5 mg), purge withhydrogen was performed for 3 times, and the reaction was performed underhydrogen (0.4 Mpa) at room temperature for 18 hours. LC-MS indicated thereaction of the starting materials was complete. The reaction solutionwas filtered, the filtrate was concentrated under reduced pressure, andthe crude product was isolated by column chromatography on silica gel(dichloromethane:methanol=10:1) to afford compound C8 (2 mg, whitesolid, yield: 13.61%, compound 8).

¹H NMR (400 MHz, CD₃OD) δ 8.39 (s, 1H), 8.25 (s, 1H), 7.60 (s, 1H), 6.7(s, 1H), 3.48-3.46 (m, 1H), 1.38 (d, J=6.4 Hz, 6H); MS m/z (ESI): 246.0[M+H]⁺.

Example 5: preparation of5-((5-isopropyl-2-methylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C9,compound 9)

Compound C2 (30 mg, 0.1 mmol) was dissolved in 1,4-dioxane (1 mL),methylboronic acid (12 mg, 0.2 mmol), K₂CO₃ (136 mg, 0.3 mmol) andPd(dppf)Cl₂ (7.5 mg, 0.01 mmol) were sequentially added, followed byaddition of purified water (0.1 mL), and purge with nitrogen wasperformed for 3 times. Under the protection of nitrogen, the reactionsolution was heated to 110° C., and the reaction was performed undermicrowave radiation for 1 hour. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was cooled toroom temperature, filtered, the filtrate was concentrated under reducedpressure to dryness, and the crude product was isolated by columnchromatography on silica gel (dichloromethane:methanol=10:1) to affordcompound C9 (3 mg, white solid, yield: 11.58%, compound 9).

¹H NMR (400 MHz, DMSO-d₆) δ 8.25 (s, 1H), 7.62 (s, 1H), 6.84 (s, 2H),6.50 (s, 1H), 6.41 (s, 2H), 3.30-3.28 (m, 1H), 2.34 (s, 3H), 1.27 (d,J=6.8 Hz, 6H); MS m/z (ESI): 259.9 [M+H]⁺.

The following compounds were prepared according to methods similar tothat described in Example 5.

Starting material or Compound regent different from No. CompoundStructure Name that in Example 5 Characterization Data C13 (compound 13)

5-(5- isopropyl-2- phenylpyridin-4- yl)oxy)pyrimidine- 2,4-diamineMethylboronic acid was replaced with phenylboronic acid. ¹H NMR (400MHz, DMSO- d₆) δ 8.50 (s, 1H), 7.84 (d, J = 7.6 Hz, 2H), 7.65 (s, 1H),7.47-7.39 (m 3H), 6.95 (s, 1H), 6.63 (s, 2H), 6.18 (s, 2H), 3.43-3.37(m, 1H), 1.34 (d, J = 7.2 Hz, 6H); MS m/z (ESI): 321.9 [M + H]⁺. C43(compound 43)

5-(5- isopropyl-2- vinylpyridin-4- yl)oxy)pyrimidine- 2,4-diamineMethylboronic acid was replaced with vinylboronic acid. ¹H NMR (400 MHz,CD₃OD) δ 8.31 (s, 1H), 7.59 (s, 1H), 6.76 (s, 1H), 6.71- 6.67 (m, 1H),6.00 (d, J = 17.6 Hz, 1H), 5.43 (d, J = 10.8 Hz, 1H), 3.45-3.41 (m, 1H),1.38 (d, J = 6.8 Hz, 6H); MS m/z (ESI): 271.9 [M + H]⁺. C44 (compound44)

5-(5- isopropyl-2- (prop-1-en-2- yl)pyridin-4- yl)oxy)pyrimidine-2,4-diamine Methylboronic acid was replaced with prop-1-en-2- ylboronicacid. ¹H NMR (400 MHz, DMSO- d₆) δ 8.38 (s, 1H), 7.60 (s, 1H), 6.68 (s,1H), 6.62 (s, 2H), 6.18 (s, 2H), 5.61 (s, 1H), 5.20 (s, 1H), 3.30-3.28(m, 1H), 2.03 (s, 3H), 1.30 (d, J = 7.2 Hz, 6H); MS m/z (ESI): 285.9[M + H]⁺. C35 (compound 35)

5-((5- isopropyl-2- (1H-pyrazol-4- yl)pyridin-4- yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with 1H-pyrazol-4- boronicacid. ¹H NMR (400 MHz, CD₃OD) δ 8.30 (s, 1H), 8.01 (s, 2H), 7.61 (s,1H), 6.86 (s, 1H), 3.45-3.43 (m, 1H), 1.39 (d, J = 6.8 Hz, 6H); MS m/z(ESI): 311.9 [M + H]⁺. C19 (compound 19)

4-((2,4- diaminopyrimidin- 5-yl)oxy)-5- isopropylpyridine- 2-thiolMethylboronic acid was replaced with triisopropylsilan ethiol;Pd(dppf)Cl₂ was replaced with Pd₂(dba)₃ and Xantphos; potassiumcarbonate was replaced with DIPEA. ¹H NMR (400 MHz, CD₃OD) δ 7.70 (s,1H), 7.55 (s, 1H), 6.71 (s, 1H), 3.30- 3.23 (m, 1H), 1.40 (d, J = 6.8Hz, 6H). MS m/z (ESI): 277.9 [M + H]⁺. C34 (compound 34)

5-((5- isopropyl-2-(1- methyl-1H- pyrazol-4- yl)pyridin-4- yl)oxy)pyrimidine- 2,4-diamine Methylboronic acid was replaced with(1-methyl-1H- pyrazol-4- yl)boronic acid; Pd(dppf)Cl₂ was replaced withPd(PPh₃)₄. ¹H NMR (400 MHz, CD₃OD) δ 8.29 (s, 1H), 8.05 (s, 1H), 7.86(s, 1H), 7.60 (s, 1H), 6.81 (s, 1H), 3.89 (s, 3H), 3.45-3.41 (m, 1H). MSm/z (ESI): 325.9 [M + H]⁺. C38 (compound 38)

5-((5- isopropyl-2- (1H-pyrrol-1- yl)pyridin-4- yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with 1H-pyrrole; potassiumcarbonate was replaced with potassium tert- butoxide; Pd(dppf)Cl₂ wasreplaced with x-phos and Pd₂(dba)₃; no water was added. ¹H NMR (400 MHz,DMSO- d₆) δ 12.24 (s, 1H), 8.48 (s, 1H), 8.29 (s, 1H), 8.09 (s, 1H),7.95 (s, 1H), 7.69-7.68 (m, 4H), 7.17 (s, 1H), 6.25 (s, 2H), 3.32-3.27(m, 1H), 1.31-1.29 (d, J = 7.2 Hz, 6H). MS m/z (ESI): 310.9 [M + H]⁺.C39 (compound 39)

5-((2-(furan-2- yl)-5- isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with furan-2- ylboronicacid; Pd(dppf)Cl₂ was replaced with Pd(PPh₃)₄. ¹H NMR (400 MHz, CD₃OD) δ8.43 (s, 1H), 7.92 (s, 1H), 7.83 (s, 1H), 7.50 (s, 1H), 7.45 (s, 1H),6.71 (s, 1H), 3.47-3.44 (m, 1H), 1.41 (d, J = 6.8 Hz, 6H). MS m/z (ESI):311.9 [M + H]⁺. C40 (compound 40)

5-((2-(1H- indol-1-yl)-5- isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with 1H-indole; potassiumcarbonate was replaced with potassium phosphate; Pd(dppf)Cl₂ wasreplaced with cyclohexane- 1,2-diamine and cuprous iodide; no water wasadded. 1H NMR (400 MHz, DMSO- d₆) δ 8.39 (s, 1H), 8.04 (d, J = 7.6 Hz,1H), 7.77 (d, J = 3.6 Hz, 1H), 7.70 (s, 1H), 7.63 (d, J = 7.6 Hz, 1H),7.22- 7.12 (m, 2H), 6.72 (s, 1H), 6.68 (d, J = 3.6 Hz, 1H), 6.58 (s,2H), 6.10 (s, 2H), 3.42- 3.33 (m, 1H), 1.36 (d, J = 7.2 Hz, 6H). MS m/z(ESI): 360.9 [M + H]⁺. C45 (compound 45)

5-((2-(1- ethoxyvinyl)-5- isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with tributyl(1-ethoxyvinyl) stannane; Pd(dppf)Cl₂ was replaced with Pd(PPh₃)₄; no waterwas added. ¹H NMR (400 MHz, CD₃OD) δ 8.32 (s, 1H), 7.57 (s, 1H), 6.97(s, 1H), 5.15 (s, 1H), 4.37 (s, 1H), 3.92-3.89 (m, 2H), 3.46-3.46 (m,1H), 1.39 (d, J = 8.0 Hz, 6H), 1.33 (d, J = 6.4 Hz, 3H). MS m/z (ESI):315.9 [M + H]⁺. C52 (compound 52)

5-((2- cyclopropyl-5- isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with cyclopropylboronicacid; potassium carbonate was replaced with potassium phosphate;Pd(dppf)Cl₂ was replaced with palladium acetate andtricyclohexylphosphine; ¹H NMR (400 MHz, DMSO- d₆) δ 12.31 (s, 1H), 8.52(s, 1H), 8.34 (s, 1H), 8.11 (s, 1H), 7.96 (s, 1H), 7.74 (s, 2H), 7.00(s, 1H), 2.34 (s, 1H), 1.27 (d, J = 6.8, 6H), 1.04-0.98 (m, 4H). MS m/z(ESI): 286.0 [M + H]⁺. the solvent, 1,4- dioxane, was replaced withtoluene; no water was added. C115 (compound 115)

5-((5- isopropyl-2-(5- methylthiazol- 2-yl)pyridin-4- yl)oxy)pyrimidine- 2,4-diamine Methylboronic acid was replaced with 5-methyl-2- (tributylstannyl) thiazole; Pd(dppf)Cl₂ was replaced withPd(PPh₃)₄. ¹H NMR (400 MHz, CD₃OD) δ 8.37 (s, 1H), 7.62 (s, 1H), 6.49(s, 1H), 7.39 (s, 1H), 3.49-3.46 (m, 1H), 2.50 (s, 3H), 1.40 (d, J = 6.8Hz, 6H). MS m/z (ESI): 342.8 [M + H]⁺. C116 (compound 116)

5-((2-(tert- butylthio)-5- isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with potassium tert-butanethiol; potassium carbonate was replaced with DIPEA; Pd(dppf)Cl₂was replaced with Xantphos and Pd₂(dba)₃. ¹H NMR (400 MHz, CD₃OD) δ 8.35(s, 1H), 7.63 (s, 1H), 6.74 (s, 1H), 3.46- 3.43 (m, 1H), 1.40-1.37 (m,15H). MS m/z (ESI): 333.9 [M + H]⁺. C117 (compound 117)

5-((2- (ethylthio)-5- isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine Methylboronic acid was replaced with sodium ethanethiolate;potassium carbonate was replaced with DIPEA; Pd(dppf)Cl₂ was replacedwith Xantphos and Pd₂(dba)₃. ¹H NMR (400 MHz, CD₃OD) δ 8.23 (s, 1H),7.60 (s, 1H), 6.48 (s, 1H), 3.40- 3.36 (m, 1H), 3.06-3.04 (m, 2H)1.38-1.29 (m, 9H). MS m/z (ESI): 305.8 [M + H]⁺. C208 (compound 208)

5-((2- (methylthio)-5- (prop-1-en-2- yl)pyridin-4- yl)oxy) pyrimidine-2,4-diamine C2 was replaced with C215-1; methylboronic acid was replacedwith sodium thiomethoxide; potassium carbonate was replaced with DIPEA;Pd(dppf)Cl₂ was replaced with X-phos and Pd₂(dba)₃; no water was added.¹H NMR (400 MHz, CD₃OD) δ 8.29 (s, 1H), 7.62 (s, 1H), 6.78 (s, 1H), 5.29(s, 1H), 5.27 (s, 1H), 2.53 (s, 3H), 2.18 (s, 3H). MS m/z (ESI): 290.0[M + H]⁺.

Example 6: preparation of4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropyl-1-methylpyridin-2(1H)-one(C11, compound 11)

Step 1:

Compound C4-2 (2.2 g, 7 mmol) was dissolved in methanol (10 mL), sodiummethoxide (610 mg, 7.7 mmol) was added, and the reaction was performedunder reflux for 16 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete. The reaction solution wasconcentrated under reduced pressure to afford a crude product, C11-1(1.9 g), which was directly used in the next step. MS m/z (ESI): 243.9[M+H]⁺.

Step 2:

In a 50 mL flask, compound C11-1 (crude product, 1.2 g, 5 mmol) wasdissolved in N,N-dimethylformamide (10 mL), potassium carbonate (1.38 g,10 mmol) was added, followed by slow dropwise addition of iodomethane(840 mg, 6 mmol), and the reaction was performed at room temperature for4 hours. LC-MS analysis indicated the reaction was complete. Thereaction was quenched by water (10 mL), then extracted with ethylacetate (20 mL×4). The organic phases were combined, washed withsaturated brine (10 mL×2), then dried over sodium sulfate, filtered, andconcentrated to give a crude product, which was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=10:1˜1:1),to afford compound C11-2 (240 mg, colorless oil, yield: 18.62%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.45 (s, 1H), 7.34-7.22 (m, 5H), 5.82 (s,1H), 5.03 (s, 2H), 3.76 (s, 3H), 2.89-2.82 (m, 1H), 1.10 (d, J=6.8 Hz,6H); MS m/z (ESI): 257.9 [M+H]⁺.

Step 3:

In a 50 mL flask, compound C11-2 (240 mg, 0.93 mmol) was dissolved inmethanol (5 mL), 10% Pd/C (20 mg) was added, purge with hydrogen wasperformed 3 times, and the reaction was performed under hydrogen at roomtemperature for 72 hours. LC-MS analysis indicated the reaction wascomplete. The reaction solution was filtered, the filter cake was rinsedwith methanol (5 mL), and the filtrate was concentrated under reducedpressure to afford compound C11-3 (150 mg, white solid, yield: 95.58%).

¹H NMR (400 MHz, DMSO-d₆) δ 6.93 (s, 1H), 5.68 (s, 1H), 3.74 (s, 3H),2.87-2.77 (m, 1H), 1.08 (d, J=6.8 Hz, 6H); MS m/z (ESI): 168.0 [M+H]⁺.

Step 4:

In a 50 mL flask, compound C11-3 (150 mg, 0.93 mmol) was dissolved inN,N-dimethylformamide (5 mL), potassium carbonate (380 mg, 1.86 mmol)was added, followed by slow dropwise addition of bromoacetonitrile (162mg, 1.41 mmol), and the reaction was performed at room temperature for18 hours. LC-MS analysis indicated the reaction was complete. Thereaction was quenched by water (10 mL), then extracted with ethylacetate (20 mL) for 4 times. The organic phases were combined, washedonce with saturated brine (10 mL), then dried over sodium sulfate,filtered, and concentrated to give a crude product. The crude productwas purified by preparative chromatography (petroleum ether:ethylacetate=3:1˜1:1) to afford compound C11-4 (110 mg, colorless oil, yield:57.42%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.48 (s, 1H), 5.88 (s, 1H), 4.91 (s, 2H),3.79 (s, 3H), 2.89-2.83 (m, 1H), 1.11 (d, J=6.8 Hz, 6H); MS m/z (ESI):207.0 [M+H]⁺.

Step 5:

In a 50 mL flask, compound C11-4 (86 mg, 0.42 mmol) was dissolved inN,N-dimethylformamide (5 mL), tert-butoxy bis(dimethylamino)methane (208mg, 1.26 mmol) was added, and the reaction solution was heated to 100°C. and allowed to proceed for 2 hours. LC-MS analysis indicated thereaction was complete. The reaction was quenched by water (10 mL), thenextracted with ethyl acetate (20 mL) for 4 times. The organic phaseswere combined, washed twice with saturated brine (10 mL), then driedover sodium sulfate, filtered, and concentrated under reduced pressure.The crude product was purified by preparative chromatography (petroleumether:ethyl acetate=3:1˜1:1) to afford compound C11-5 (60 mg, colorlessoil, yield: 46.69%). MS m/z (ESI): 307.0 [M+H]⁺.

Step 6:

In a 50 mL flask, compound C11-5 (60 mg, 0.2 mmol) was dissolved inN,N-dimethylformamide (2 mL), aniline hydrochloride (32 mg, 0.24 mmol)was added, the reaction was heated to 100° C., and stirred for 2 hours.LC-MS indicated the reaction of the starting materials was complete. Thereaction was added with 25 mL water, and extracted with ethyl acetate(20 mL×3). The organic phase was washed twice with saturated brine (20mL), dried over anhydrous sodium sulfate for half an hour, and filtered.The filtrate was concentrated under reduced pressure, and the crudeproduct was purified by preparative chromatography (petroleumether:ethyl acetate=2:1) to afford compound C11-6 (30 mg, colorless oil,yield: 48.54%).

¹H NMR (400 MHz, DMSO-d₆) δ 9.23 (d, J=13.2 Hz, 1H), 8.00 (d, J=13.2 Hz,1H), 7.37-7.21 (m, 4H), 7.09 (s, 1H), 6.99 (d, J=6.8 Hz, 1H), 5.91 (s,1H), 3.82 (s, 3H), 2.89-2.82 (m, 1H), 1.13 (d, J=6.8 Hz, 6H); MS m/z(ESI): 309.9 [M+H]⁺.

Step 7:

In a 50 mL flask, compound C11-6 (20 mg, 0.07 mmol) was dissolved inanhydrous ethanol (2 mL), guanidine carbonate (38 mg, 0.21 mmol) wasadded, the reaction was heated to 100° C. under microwave radiation, andstirred for 2 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was cooled to room temperature, andconcentrated under reduced pressure. The crude product was purified bypreparative chromatography (dichloromethane:methanol=10:1) to affordcompound C11 (5 mg, white solid, yield: 25.97%, compound 11).

¹H NMR (400 MHz, DMSO-d₆) δ 7.52 (s, 1H), 6.98 (s, 1H), 6.12 (s, 4H),5.85 (s, 1H), 3.80 (s, 3H), 2.83-2.91 (m, 1H), 1.12 (d, J=6.8 Hz, 6H);MS m/z (ESI): 275.9 [M+H]⁺.

Example 7: preparation of4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropyl-2-methoxypyridine 1-oxide(C14, compound 14)

Compound C4 (10 mg, 0.036 mmol) was dissolved in anhydrousdichloromethane (5 mL), followed by addition of meta-chloroperbenzoicacid (13 mg, 0.072 mmol), and the reaction was performed at roomtemperature for 18 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction solution was filtered, the filtratewas concentrated under reduced pressure, and the crude product wasseparated by thin layer chromatography (dichloromethane:methanol=10:1)to afford compound C14 (2 mg, white solid, yield: 19.16%, compound 14).

¹H NMR (400 MHz, CD₃OD) δ 7.95 (s, 1H), 7.59 (s, 1H), 6.02 (s, 1H), 3.82(s, 3H), 3.31-3.28 (m, 1H), 1.35 (d, J=6.8 Hz, 6H); MS m/z (ESI): 291.9[M+H]⁺.

The following compound was prepared according to a method similar tothat described in Example 7.

Starting material or regent different Compound from that in No CompoundStructure Name Example 7 Characterization Data C1 (compound 1)

2-bromo-4- ((2,4- diaminopyrimidin- 5-yl)oxy)-5- isopropylpyridine1-oxide C4 was replaced with C2. ¹H NMR (400 MHz, CD₃OD) δ 8.19 (s, 1H),7.65 (s, 1H), 6.88 (s, 1H), 3.45-3.38 (m, 1H), 1.37 (d, J = 6.8 Hz, 6H).MS m/z (ESI): 339.8 [M + H]⁺.

Example 8: preparation of5-((2-ethyl-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C15,compound 15)

Compound C43 (4 mg, 0.014 mmol) was dissolved in anhydrous methanol (5mL), followed by addition of 10% palladium/carbon (4 mg), purge withhydrogen was performed for 3 times, and the reaction was performed underhydrogen (0.4 Mpa) at room temperature for 18 hours. LC-MS indicated thereaction was complete. The reaction solution was filtered, the filtratewas concentrated under reduced pressure, and the crude product wasisolated by thin layer chromatography (dichloromethane:methanol=10:1) toafford compound C15 (2 mg, white solid, yield: 52.33%, compound 15).

¹H NMR (400 MHz, CD₃OD) δ 8.25 (s, 1H), 7.58 (s, 1H), 6.56 (s, 1H),3.44-3.37 (m, 1H), 2.68 (q, J=7.6 Hz, 2H), 1.36 (d, J=7.2 Hz, 6H), 1.19(t, J=7.6 Hz, 3H); MS m/z (ESI): 274.0 [M+H]⁺.

Example 9: preparation of5-((2,5-diisopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C16,compound 16)

Compound C44 (4 mg, 0.014 mmol) was dissolved in anhydrous methanol (5mL), followed by addition of 10% palladium/carbon (4 mg), purge withhydrogen was performed for 3 times, and the reaction was performed underhydrogen (0.4 Mpa) at room temperature for 18 hours. LC-MS indicated thereaction of the starting materials was complete. The reaction solutionwas filtered, the filtrate was concentrated under reduced pressure, andthe crude product was separated by thin layer chromatography(dichloromethane:methanol=10:1) to afford compound C16 (2 mg, whitesolid, yield: 49.78%).

¹H NMR (400 MHz, CD₃OD) δ 8.26 (s, 1H), 7.58 (s, 1H), 6.57 (s, 1H),3.46-3.39 (m, 1H), 3.31-3.29 (m, 1H), 1.36 (d, J=7.2 Hz, 6H), 1.20 (d,J=6.8 Hz, 6H); MS m/z (ESI): 288.0 [M+H]⁺.

Example 10: preparation of(4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpyridin-2-yl)diethylphosphineoxide (C18, compound 18)

At 25° C. and under the protection of nitrogen, Pd(dppf)₂Cl₂ (10 mg) wasadded to a mixed solution of C2 (20 mg, 0.062 mmol), diethylphosphineoxide (65.72 mg, 0.62 mmol), DIEA (80 mg, 0.62 mmol) and DMF (5 mL). thereaction was stirred at 100° C. overnight. The reaction solution wasconcentrated to give a black oil, which was purified by preparativehigh-performance liquid chromatography to afford C18 (2.19 mg, whitesolid, yield: 10.12%, compound 18).

¹H NMR (400 MHz, CD₃OD) δ 8.71 (s, 1H), 7.84 (s, 1), 7.46 (d, J=6 Hz,1H), 3.55-3.52 (m, 1H), 2.13-2.04 (m, 4H), 1.43 (d, J=6.8 Hz, 6H),1.08-1.02 (m, 6H); MS m/z (ESI): 349.9 [M+H]⁺.

Example 11: preparation of5-((5-isopropyl-2-(methylthio)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C20, compound 20),5-((5-isopropyl-2-(methylsulfonyl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C5, compound 5) and5-((5-isopropyl-2-(methylsulfinyl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C48, compound 48)

Step 1:

At 25° C. and under the protection of nitrogen, Pd₂(dba)₃ (5 mg) wasadded to a mixed solution of C2 (15 mg, 0.046 mmol), sodiumthiomethoxide (20 mg, 0.285 mmol), DIEA (20 mg, 0.188 mmol) and Xantphos(15 mg, 0.026 mmol) (dissolved in 1,4-dioxane (10 mL)). The reaction wasstirred at 100° C. overnight. The reaction was concentrated to drynessto give a black oil, and the residue was purified by preparative thinlayer chromatography (DCM:MeOH=20:1) to afford compound C20 (13 mg,white solid, yield: 97.12%, compound 20).

¹H NMR (400 MHz, CD₃OD) δ 8.22 (s, 1H), 7.60 (s, 1H), 6.47 (s, 1H),3.40-3.33 (m, 1H), 2.46 (s, 3H), 1.38-1.36 (d, J=7.2 Hz, 1H); MS m/z(ESI): 291.8 [M+H]⁺.

Step 2:

At 0° C., Oxone (42.4 mg, 0.069 mmol) was added to a mixed solution ofC20 (10 mg, 0.034 mmol) in acetone (4 mL) and water (1 mL). The reactionwas stirred at 0° C. for 1 h. The reaction was quenched by water, andthen extracted with ethyl acetate. The organic phase was washed withsaturated brine, dried over anhydrous sodium sulfate, filtered, andconcentrated under reduced pressure to dryness to give a yellow residue.The residue was purified by preparative thin layer chromatography(DCM:MeOH=20:1) to afford compound C5 (4 mg, white solid, yield: 36.42%,compound 5).

¹H NMR (400 MHz, DMSO-d₆) δ 8.62 (s, 1H), 7.69 (s, 1H), 7.05 (s, 1H),6.66 (s, 2H), 6.20 (s, 2H), 3.48-3.44 (m, 1H), 3.24 (s, 3H), 1.35-1.34(d, J=6.8 Hz, 6H); MS m/z (ESI): 323.8 [M+H]⁺.

Compound C20 (20 mg, 68.6 mmol) was dissolved in acetone (5 mL), anaqueous solution (1 mL) of oxone (30 mg, 84.4 mmol) was added at 0° C.,and the reaction solution was stirred at 0° C. for 0.5 hour. Thereaction was added with water (5 mL), extracted with dichloromethane (5mL×3). The organic phase was dried over sodium sulfate, concentratedunder reduced pressure to afford a crude product. The crude product waspurified by preparative thin layer chromatography on silica gel(dichloromethane:methanol=20:1) to afford compound C48 (10 mg, whitesolid, yield 47.5%).

¹H NMR (400 MHz, CD₃OD) δ 8.53 (s, 1H), 7.67 (s, 1H), 7.25 (s, 1H),3.56-3.52 (m, 1H), 1.43 (d, J=6.8 Hz, 6H). MS m/z (ESI): 307.9 [M+H]⁺.

Example 12: preparation of5-((5-isopropyl-2-(pyrrolidin-1-yl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C63, compound 63)

At 25° C., a solution of C2 (10 mg, 0.031 mmol) in pyrrolidine (1 mL)was stirred at 120° C. (microwave) for 2 hours. The reaction wasconcentrated to dryness to give a yellow oil, which was purified bypreparative thin layer chromatography (DCM:MeOH=20:1) to afford C63(1.62 mg, light yellow solid, yield: 16.64%, compound 63).

¹H NMR (400 MHz, CD₃OD) δ 7.72 (s, 1H), 7.65 (s, 1H), 5.85 (s, 1H),3.42-3.38 (m, 4H), 3.33-3.27 (m, 1H), 2.07-2.04 (m, 4H), 1.35-1.33 (d,J=7.2 Hz, 6H); MS m/z (ESI): 314.9 [M+H]⁺.

Example 13: preparation of5-((2-(dimethylamino)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C23, compound 23)

At 25° C., dimethylamine hydrochloride (20 mg) was added to a solutionof C2 (10 mg, 0.031 mmol) in DIEA (1 mL). The reaction was stirred at120° C. (microwave) for 5 hours. The reaction solution was concentratedto dryness to give a yellow oil, which was purified by preparative thinlayer chromatography to afford compound C23 (2 mg, light yellow solid,yield: 22.40%, compound 23).

¹H NMR (400 MHz, CD₃OD) δ 7.947 (s, 1H), 7.686 (s, 1H), 6.390 (s, 1H),3.330-3.233 (m, 1H), 3.218 (s, 6H), 1.340 (d, J=6.8 Hz, 6H); MS m/z:288.9 [M+H]⁺.

Example 14: preparation of5-((2-(benzylsulfonyl)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C42, compound 42) and5-((2-(benzylthio)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C41, compound 41)

Step 1:

At 25° C. and under the protection of nitrogen, Pd₂(dba)₃ (10 mg) wasadded to a mixed system of C2 (25 mg, 0.077 mmol), benzyl mercaptan (19mg, 015 mmol), DIEA (24.6 mg, 0.23 mmol), Xantphos (19 mg, 0.033 mmol)and dioxane (10 mL). The reaction solution was stirred at 100° C.overnight. The reaction solution was concentrated to dryness to give ayellow oil, which was purified by preparative thin layer chromatographyon silica gel (DCM:MeOH=15:1) to afford compound C41 (12 mg, whitesolid, yield: 42.27%, compound 41).

¹H NMR (400 MHz, CD₃OD) δ 8.20 (s, 1H), 7.49 (s, 1H), 7.28-7.21 (m, 5H),6.44 (s, 1H), 4.24 (s, 2H), 2.98-2.94 (q, J=6.8 Hz, 1H), 1.36 (d, J=6.8Hz, 6H); MS m/z (ESI): 367.8 [M+H]⁺.

Step 2:

At 25° C., m-CPBA (8.3 mg) was added to a solution of C41 (8 mg, 0.0217mmol) in dichloromethane (10 mL). The reaction was stirred at 25° C. for1 h. The reaction was concentrated to dryness to give a yellow oil,which was purified by preparative thin layer chromatography(DCM:MeOH=10:1) to afford C42 (1 mg, white solid, yield: 11.55%,compound 42).

¹H NMR (400 MHz, CD₃OD) δ 8.64 (s, 1H), 7.40 (s, 1H), 7.33-7.24 (m, 3H),7.14-7.13 (m, 2H), 6.88 (s, 1H), 4.62 (s, 2H), 3.54-3.51 (q, J=6.8 Hz,1H), 1.43 (d, J=6.8 Hz, 6H); MS m/z (ESI): 399.8 [M+H]⁺.

Example 15: preparation of4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpyridine-2-sulfonamide(C6, compound 6)

C41 (0.1 g, 0.27 mmol) was dissolved in dichloromethane (5 mL) andacetic acid (5 mL), and NCS (3.0 g, 0.027 mol) was added. The reactionwas stirred at room temperature for 1 hour. The reaction solution waspoured into aqueous ammonia (10 mL), stirred for half an hour, and thenextracted with ethyl acetate. The organic phase was concentrated todryness to give a crude product, which was purified by Prep-HPLC toafford C6 (10 mg, white solid, yield 11.3%).

¹H NMR (400 MHz, CD₃OD) δ 8.60 (s, 1H), 7.85 (s, 1H), 7.37 (s, 1H),3.53-3.50 (m, 1H), 1.42 (d, J=7.2 Hz, 6H). MS m/z (ESI): 324.7 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 15.

Starting material or regent different Compound from that in No. CompoundStructure Name Example 15 Characterization Data C26 (compound 26)

4-((2,4- diaminopyrimidin- 5-yl)oxy)-5- isopropyl-N- methylpyridine-2-sulfonamide The aqueous ammonia was replaced with a solution ofmethylamine in tetrahydrofuran. ¹H NMR (400 MHz, CD₃OD) δ 8.62 (s, 1H),7.86 (s, 1H), 7.33 (s, 1H), 3.52-3.48 (m, 1H), 2.65 (s, 3H), 1.42 (d, J= 7.2 Hz, 6H). MS m/z (ESI): 338.9 [M + H]⁺ . C27 (compound 27)

4-((2,4- diaminopyrimidin- 5-yl)oxy)-5- isopropyl-N,N- dimethylpyridine-2-sulfonamide The aqueous ammonia was replaced with a solution ofdimethylamine in tetrahydrofuran. ¹H NMR (400 MHz, CD₃OD) δ 8.64 (s,1H), 7.88 (s, 1H), 7.30 (s, 1H), 3.54-3.51 (m, 1H), 2.89 (s, 6H), 1.43(d, J = 6.8 Hz, 6H). MS m/z (ESI): 352.9 [M + H]⁺. C47 (compound 47)

5-((2- (benzylsulfinyl)- 5-isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine It is a by-product isolated during the synthesis of Example15. ¹H NMR (400 MHz, DMSO- d₆) δ 7.36-7.23 (m, 6H), 7.00 (s, 1H), 6.66(s, 1H), 6.41 (s, 2H), 6.05 (s, 2H), 5.33 (s, 2H), 3.42-3.38 (m, 1H),1.33 (m, 6H). MS m/z (ESI): 383.8 [M + H]⁺.

Example 16: preparation of(4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpyridin-2-yl)dimethylphosphineoxide (C17, compound 17)

At 25° C. and under the protection of nitrogen, Pd (dppf)₂Cl₂ (10 mg)was added to a mixed solution of C2 (20 mg, 0.062 mmol),dimethylphosphine oxide (15 mg, 0.186 mmol), DIEA (24 mg, 0.186 mmol)and DMF (5 mL). The reaction was stirred at 100° C. overnight. Thereaction solution was concentrated to give a black oil, and the residuewas purified by preparative high-performance liquid chromatography toafford compound C17 (3 mg, white solid, 15.1%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.67 (s, 1H), 8.60 (s, 1H), 8.23 (s, 1H),8.03 (s, 1H), 7.81 (s, 2H), 7.25 (d, J=6 Hz, 1H), 3.45-3.38 (m, 1H),1.65 (s, 3H), 1.61 (s, 3H), 1.33 (d, J=6.8 Hz, 6H). MS m/z (ESI): 321.8[M+H]⁺.

The compound in the following table was prepared according to methodssimilar to that described in Example 16.

Starting material or regent different Compound from that in No. CompoundStructure Name Example 16 Characterization Data C49 (compound 49)

1-(4-((2,4- diaminopyrimidin- 5-yl)oxy)-5- isopropylpyridin-2-yl)phospholane 1-oxide The dimethylphosphine oxide was replaced withphospholane 1- oxide. ¹H NMR (400 MHz, DMSO-d₆) δ 8.61 (s, 1H), 7.63 (s,1H), 7.11 (d, J = 6.4 Hz, 1H), 6.55 (s, 2H), 6.12 (s, 2H), 3.44-3.40 (m,1H), 2.00-1.48 (m, 8H), 1.32 (d, J = 7.2 Hz, 6H). MS m/z (ESI): 347.8[M + H]⁺.

Example 17: preparation of5-((2-amino-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C21,compound 21)

Step 1:

Compound C21-1 (28 g, 0.138 mol) was dissolved in 1,4-dioxane (300 mL),isopropenylpinacol borate (46.3 g, 0.275 mol), potassium carbonate (38.6g, 0.276 mol), Pd(PPh₃)₄ (3.5 g) and water (30 mL) were added. Purgewith nitrogen was performed for 3 times, and the reaction solution wasstirred at 100° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was cooled toroom temperature, quenched by adding water (300 mL), and extracted withethyl acetate (300 mL×3). The organic phases were combined, washed withsaturated brine (100 mL), added with anhydrous sodium sulfate (50 g),dried for half an hour, and filtered. The filtrate was concentratedunder reduced pressure to afford a crude product, which was purified bycolumn chromatography on silica gel (petroleum ether:ethylacetate=20:1˜5:1) to afford compound C21-2 (14 g, yellow solid, yield61.9%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.63 (s, 1H), 6.01 (s, 1H), 5.88 (s, 2H),5.01-4.95 (m, 2H), 3.74-3.70 (m, 3H), 1.99 (s, 3H). MS m/z (ESI): 165.0[M+H]⁺.

Step 2:

Compound C21-2 (14 g, 0.085 mol) was dissolved in ethanol (200 mL), 10%wet palladium/carbon (10 g) was added, and the reaction solution wasstirred under hydrogen at room temperature overnight. LC-MS indicatedthe reaction of the starting materials was substantially complete. Thereaction was filtered, concentrated under reduced pressure to afford acrude product, which was purified by column chromatography on silica gel(dichloromethane:methanol=20:1) to afford compound C21-3 (12 g, whitesolid, yield 84.5%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.58 (s, 1H), 5.99 (s, 1H), 5.61 (s, 2H),3.73 (s, 3H), 2.94-2.93 (m, 1H), 1.16-1.08 (m, 6H). MS m/z (ESI): 167.0[M+H]⁺.

Step 3:

Compound C21-3 (5 g, 0.030 mol) was dissolved in dichloromethane (20mL), boron tribromide (10 mL) was added, the reaction solution wasstirred at 40° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was slowlydropwise added to ice water, adjusted to pH 8 with sodium carbonate, andextracted with ethyl acetate (100 mL×3). The organic phases werecombined, washed once with saturated brine (100 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated to give a crude product, C21-4, and the crude product wasdirectly used in the next step (4 g, crude product, yellow oily liquid,yield 89%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.23 (s, 1H), 7.13 (s, 1H), 6.55 (s, 1H),5.77-5.43 (m, 2H), 2.85 (m, 1H), 1.13-1.05 (m, 6H). MS m/z (ESI): 153.0[M+H]⁺.

Step 4:

Compound C21-4 (3 g, 0.020 mol) was dissolved in acetonitrile (30 mL),di-tert-butyl dicarbonate (8.6 g, 0.039 mol), triethylamine (3.9 g,0.039 mol) and DMAP (2.4 g, 0.019 mol) were sequentially added, and thereaction solution was stirred at 30° C. for 3 hours. LC-MS indicated thereaction of the starting materials was substantially complete. Thereaction was quenched by adding water (100 mL), and extracted with ethylacetate (100 mL×3). The organic phases were combined, washed once withsaturated brine (100 mL), then dried over anhydrous sodium sulfate (10g) for half an hour, filtered, and concentrated under reduced pressureto afford a crude product, which was purified by column chromatographyon silica gel (petroleum ether:ethyl acetate=10:1) to afford compoundC21-5 (1.4 g, white solid, yield 29%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.48 (s, 1H), 9.39 (s, 1H), 7.84 (s, 1H),7.35 (s, 1H), 3.02 (s, 1H), 1.47 (s, 9H), 1.17 (s, 6H). MS m/z (ESI):253.1 [M+H]⁺.

Step 5:

Compound C21-5 (1.4 g, 5.56 mmol) was dissolved in DMF (15 mL),bromoacetonitrile (1 g, 8.30 mmol) and potassium carbonate (1.5 g, 11.10mmol) were sequentially added, and the reaction solution was stirred atroom temperature for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction was quenched by addingwater (100 mL), and extracted with ethyl acetate (100 mL×3). The organicphases were combined, washed once with saturated brine (100 mL), thendried over anhydrous sodium sulfate (10 g) for half an hour, filtered,and concentrated under reduced pressure to afford a crude product, whichwas purified by column chromatography on silica gel (petroleumether:ethyl acetate=5:1˜1:1) to afford compound C21-6 (790 mg, yellowsolid, yield 38%).

¹H NMR (400 MHz, DMSO-d₆) δ 9.77 (s, 1H), 8.02 (s, 1H), 7.53 (s, 1H),5.27 (s, 2H), 3.06-3.02 (m, 1H), 1.48 (s, 9H), 1.20-1.19 (m, 6H). MS m/z(ESI): 291.9 [M+H]⁺.

Step 6:

Compound C21-6 (300 mg, 1.03 mmol) was dissolved in DMF (5 mL),tert-butoxy bis(dimethylamino)methane (718 mg, 4.12 mmol) was added, andthe reaction solution was stirred at 100° C. for 1 hour. LC-MS indicatedthe reaction of the starting materials was complete. The reactionsolution was concentrated under reduced pressure, to afford a crudeproduct, C21-7, and the crude product was directly used in the next step(0.4 g, yellow oily liquid, yield 99%). MS m/z (ESI): 318.0 [M+H]⁺.

Step 7:

Compound C21-7 (400 mg, 1.03 mmol) was dissolved in DMF (5 mL), anilinehydrobromide (285 mg, 3.07 mmol) was added, and the reaction solutionwas stirred at 100° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was cooled toroom temperature, quenched by adding water (30 mL), and extracted withethyl acetate (50 mL×3). The organic phases were combined, washed withsaturated brine (50 mL), added with anhydrous sodium sulfate (10 g),dried for half an hour, and filtered. The filtrate was concentratedunder reduced pressure to afford a crude product, which was purified bycolumn chromatography on silica gel (petroleum ether:ethylacetate=5:1˜1:1) to afford compound C21-8, (230 mg, yellow oily liquid,yield 61%). MS m/z (ESI): 365.9 [M+H]⁺.

Step 8:

Compound C21-8 (120 mg, 0.33 mmol) was dissolved in ethanol (5 mL),guanidine hydrochloride (150 mg, 0.99 mmol) and sodium methoxide (53 mg,0.99 mmol) were sequentially added, and the reaction solution wasstirred at 90° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was cooled toroom temperature, concentrated under reduced pressure to afford a crudeproduct, C21-9, and the crude product was directly used in the next step(180 mg, brown oily liquid, yield 100%). MS m/z (ESI): 331.8 [M+H]⁺.

Step 9:

Compound C21-9 (90 mg, 0.27 mmol) was dissolved in ethanol (5 mL), anaqueous solution of sodium hydroxide (109 mg, 2.7 mmol, 1 mL H₂O) wasadded, and the reaction was stirred at 90° C. for 6 hours. LC-MSindicated the reaction of the starting materials was complete. Thereaction solution was cooled to room temperature, concentrated underreduced pressure, and the crude product was isolated by preparativeliquid chromatography to afford compound C21 (15 mg, white solid, yield21%).

¹H NMR (400 MHz, CD₃OD) δ 7.96 (s, 1H), 7.70 (s, 1H), 6.33 (s, 1H),3.30-3.28 (m, 1H), 1.34-1.30 (d, J=7.2 Hz, 6H). MS m/z (ESI): 260.9[M+H]⁺.

Example 18: preparation of5-((5-isopropyl-2-(methylamino)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C22, compound 22)

Step 1:

Compound C22-1 (3 g, 0.018 mol) was dissolved in tert-butanol (200 mL),di-tert-butyl dicarbonate (4.73 g, 0.021 mol) was added, and thereaction solution was stirred at 20° C. for 16 hours. LC-MS indicatedthe reaction of the starting materials was substantially complete. Thereaction solution was concentrated under reduced pressure to afford acrude product, which was purified by column chromatography on silica gel(petroleum ether:ethyl acetate=100:1˜50:1) to afford compound C22-2 (2.5g, white solid, yield 52%).

¹H NMR (400 MHz, DMSO-d₆) δ 9.72 (s, 1H), 7.92 (s, 1H), 7.45 (s, 1H),3.84 (s, 3H), 3.07-3.05 (m, 1H), 1.48 (s, 9H), 1.18 (d, J=7.2 Hz, 6H).MS m/z (ESI): 267.0 [M+H]⁺.

Step 2:

Compound C22-2 (2.5 g, 0.094 mol) was dissolved in DMF (25 mL), sodiumhydride (451 mg, 18.8 mmol) was added, and the reaction solution wasstirred at 0° C. for 15 minutes. Iodomethane (2.0 g, 14.1 mmol) was thenadded, and the reaction solution was stirred at 0° C. for 1 hour. LC-MSindicated the reaction of the starting materials was substantiallycomplete. Then, the reaction was added with purified water (150 mL), andextracted with ethyl acetate (200 mL×3). The organic phases werecombined, washed with an aqueous solution of NaCl (100 mL), added withanhydrous sodium sulfate (20 g), dried for 30 min, filtered, andconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=30:1˜20:1) to afford compound C22-3 (2.2 g, brown oily liquid,yield 84%). MS m/z (ESI): 280.9 [M+H]⁺.

Step 3:

Compound C22-3 (1.2 g, 0.043 mol) was dissolved in dichloromethane (10mL), boron tribromide (10 mL) was added, and the reaction solution wasstirred at 40° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was slowlydropwise added to ice water, adjusted to pH 8 with sodium carbonate, andextracted with ethyl acetate (100 mL×3). The organic phases werecombined, washed once with saturated brine (100 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated to give a crude product, C22-4, and the crude product wasdirectly used in the next step (1 g crude product, yellow oily liquid,yield 80%). MS m/z (ESI): 167.0 [M+H]⁺.

Step 4:

Compound C22-4 (1 g, 6.02 mmol) was dissolved in DMF (10 mL),bromoacetonitrile (723 mg, 6.02 mmol) and sodium carbonate (1.92 g,18.06 mmol) were sequentially added, and the reaction solution wasstirred at room temperature for 16 hours. LC-MS indicated the reactionof the starting materials was complete. The reaction was quenched byadding water (100 mL), and extracted with ethyl acetate (100 mL×3). Theorganic phases were combined, washed once with saturated brine (100 mL),then dried over anhydrous sodium sulfate (10 g) for half an hour,filtered, and concentrated under reduced pressure to afford a crudeproduct, which was purified by column chromatography on silica gel(petroleum ether:ethyl acetate=5:1˜1:1) to afford compound C22-5 (0.17g, yellow solid, yield 15%).

¹H NMR (400 MHz, DMSO-d₆) δ 7.73 (s, 1H), 6.11 (s, 1H), 5.22 (s, 2H),2.93-2.95 (m, 1H), 2.77 (s, 3H), 1.16 (d, J=6.8 Hz, 6H). MS m/z (ESI):206.0 [M+H]⁺.

Step 5:

Compound C22-5 (170 mg, 0.83 mmol) was dissolved in DMF (2 mL),tert-butoxy bis(dimethylamino)methane (577 mg, 3.32 mmol) was added, andthe reaction solution was stirred at 100° C. for 1 hour. LC-MS indicatedthe reaction of the starting materials was complete. The reactionsolution was concentrated under reduced pressure to afford a crudeproduct, C22-6, and the crude product was directly used in the next step(0.2 g, yellow oily liquid, yield 79%). MS m/z (ESI): 260.9 [M−45+H]*.

Step 6:

Compound C22-6 (100 mg, 0.33 mmol) was dissolved in DMF (2 mL), anilinehydrobromide (54 mg, 0.39 mmol) was added, and the reaction solution wasstirred at 100° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was cooled toroom temperature, quenched by adding water (30 mL), and extracted withethyl acetate (50 mL×3). The organic phases were combined, washed withsaturated brine (50 mL), added with anhydrous sodium sulfate (10 g),dried for half an hour, and filtered. The filtrate was concentratedunder reduced pressure to afford a crude product, C22-7, and the crudeproduct was directly used in the next step (70 mg, brown oily liquid,yield 70%). MS m/z (ESI): 308.9 [M+H]⁺.

Step 7:

Compound C22-7 (70 mg, 0.23 mmol) was dissolved in ethanol (15 mL),guanidine hydrochloride (65 mg, 0.68 mmol) and sodium methoxide (37 mg,0.68 mmol) were sequentially added, and the reaction was stirred at 90°C. for 16 hours. LC-MS indicated the reaction of the starting materialswas complete. The reaction solution was cooled to room temperature,concentrated under reduced pressure, and the crude product was isolatedby preparative liquid chromatography to afford compound C22 (12 mg,light yellow solid, yield 19%).

¹H NMR (400 MHz, CD₃OD) δ 7.97 (s, 1H), 7.64 (s, 1H), 6.32 (s, 1H),3.30-3.28 (m, 1H), 2.99 (s, 3H), 1.34 (d, J=6.8 Hz, 6H). MS m/z (ESI):274.9 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 18.

Starting material or regent different Compound from that in No. CompoundStructure Name Example 18 Characterization Data C120 (compound 120)

5-((5- isopropyl-2- (isopropylamino) pyridin-4- yl)oxy)pyrimidine-2,4-diamine Iodomethane in Step 2 was replaced with isopropyl iodine. ¹HNMR (400 MHz, CD₃OD) δ 7.98 (s, 1H), 7.61 (s, 1H), 6.27 (s, 1H),3.88-3.84 (m, 1H), 3.33-3.32 (m, 1H), 1.34 (d, J = 6.8 Hz, 6H), 1.29 (d,J = 6.4 Hz, 6H). MS m/z (ESI): 302.9 [M + H]⁺. C118 (compound 118)

5-((2- (ethylamino)- 5-isopropylpyridin- 4-yl)oxy) pyrimidine-2,4-diamine C118-1 was prepared from C22-1^(i), and the preparationstarted from Step 3 of Example 18. ¹H NMR (400 MHz, CD₃OD) δ 7.74 (s,1H), 7.56 (s, 1H), 5.77 (s, 1H), 3.26-3.17 (m, 3H), 1.33 (d, J = 6.8 Hz,6H), 1.17-1.14 (m, 3H). MS m/z (ESI): 288.8 [M + H]⁺. C119 (compound119)

5-((2- (cyclopropylamino)- 5-isopropylpyridin- 4-yl)oxy)pyrimidine-2,4-diamine Step 2 of Example 18 was replaced with the preparation ofC119-1^(iii). ¹H NMR (400 MHz, CD₃OD) δ 7.79 (s, 1H), 7.58 (s, 1H), 5.99(s, 1H), 3.37 (s, 1H), 2.23- 2.19 (m, 1H), 1.34- 1.31 (m, 6H), 1.02-0.94 (m, 2H), 0.64- 0.62 (m, 1H), 0.38- 0.37 (m, 1H). MS m/z (ESI):301.0 [M + H]⁺. C61 (compound 61)

5-((2- (ethyl(methyl) amino)-5- isopropylpyridin- 4-yl)oxy)pyrimidine-2,4-diamine The preparation started from Step 3 of Example 18, and C22-3in Step 3 was replaced with C61-1^(ii). ¹H NMR (400 MHz, DMSO-d₆) δ 7.88(s, 1H), 7.52 (s, 1H), 6.32 (s, 2H), 5.98 (s, 2H), 5.62 (s, 1H),3.42-3.40 (m, 2H), 3.17-3.16 (m, 1H), 2.80 (s, 3H), 1.23 (d, J = 6.8 Hz,6H), 0.98-0.95 (m, 3H). MS m/z (ESI): 303.0 [M + H]⁺. C65 (compound 65)

5-((2- (cyclopropyl (methyl)amino)- 5-isopropylpyridin- 4-yl)oxy)pyrimidine- 2,4-diamine The preparation started from Step 3 of Example18, and C22-3 in Step 3 was replaced with C65-5^(ii). ¹H NMR (400 MHz,CD₃OD) δ 7.99 (s, 1H), 7.76 (s, 1H), 6.53 (s, 1H), 3.21 (s, 3H), 2.91-2.81 (m, 1H), 2.25- 2.18 (m, 1H), 1.35 (d, J = 7.0 Hz, 6H), 1.09- 1.04(m, 2H), 0.84- 0.80 (m, 2H). MS m/z (ESI): 315.1 [M + H]⁺. C152(compound 152)

5-((5- isopropyl-2- (2,2,2- trifluoroethyl) amino) pyridin-4- yl)oxy)pyrimidine- 2,4-diamine The preparation started from Step 3 of Example18, and C22-3 in Step 3 was replaced with C152-3^(iv). ¹H NMR (400 MHz,DMSO-d₆) δ 7.81 (s, 1H), 7.54 (s, 1H), 6.91 (t, J = 6.4 Hz, 1H), 6.31(s, 2H), 5.99 (s, 2H), 5.84 (s, 1H), 4.09-4.02 (m, 2H), 3.20-3.16 (m,1H), 1.24 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 343.0 [M + H]⁺.

i Preparation of Intermediate Example C118-1

Compound C22-1 (10 g, 0.060 mol) was dissolved in ethanol (30 mL),acetaldehyde (2.7 g, 0.06 mol) was added, then sodium borohydride (6.9g, 0.18 mol) was slowly added, and the reaction solution was stirred at25° C. for 4 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete. The reaction solution was quenchedby adding water (100 mL), and extracted with ethyl acetate (120 mL×3).The organic phases were combined, washed with saturated brine (100 mL),added with anhydrous sodium sulfate (10 g), dried for half an hour, andfiltered. The reaction solution was concentrated under reduced pressureto afford a crude product, which was purified by column chromatographyon silica gel (petroleum ether:ethyl acetate=10:1˜5:1) to affordcompound C118-1 (8.5 g, yellow solid, yield 52%). MS m/z (ESI): 195.0[M+H]⁺.

ii Preparation of Intermediate Example C65-5 and C61-1

Step 1:

To a solution of C22-1 (10 g, 0.06 mol) in dichloromethane (200 mL), anaqueous solution of formaldehyde (10 mL, 0.12 mol) and anhydrousmagnesium sulfate (30 g) as well as several drops of acetic acid wereadded, and the reaction was stirred at room temperature for 1 h. Thereaction solution was added with sodium borohydride (12 g, 0.31 mol),and then stirred at room temperature for 18 h. The reaction wascomplete. The reaction was filtered, the filter cake was washed withdichloromethane (100 mL), and the filtrate was sequentially washed withwater (200 mL) and saturated brine (200 mL×2). The organic phase wasdried over anhydrous sodium sulfate, filtered, and concentrated underreduced pressure. The residue was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=1:3) to afford C65-2 (2.1 g,light yellow oil, yield 19.4%).

¹H NMR (300 MHz, CDCl₃) δ 7.79 (s, 1H), 5.83 (s, 1H), 3.83 (s, 3H),3.11-3.02 (m, 1H), 2.92 (d, J=5.0 Hz, 3H), 1.21 (d, J=6.9 Hz, 6H). MSm/z (ESI): 181.2 [M+H]⁺.

Step 2:

C65-2 (1.9 g, 11 mmol) was dissolved in dichloromethane (40 mL), borontribromide (10 g, 40 mmol) was dropwise added at room temperature, afterthe dropwise addition, the reaction was placed in an oil bath at 50° C.,and allowed to proceed for 18 h. After the reaction was complete, thereaction system was quenched by adding methanol (50 mL) under ice bathcooling, and concentrated under reduced pressure. The residue wasadjusted to pH 6˜7 with a saturated aqueous solution of sodiumbicarbonate, extracted with tetrahydrofuran (50 mL×8), the combinedorganic phase was dried over anhydrous sodium sulfate, filtered, andthen the filtrate was concentrated under reduced pressure, to affordC65-3 (1.2 g, light yellow solid, yield 68.5%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.90 (s, 1H), 7.35 (s, 1H), 6.52 (s, 1H),5.72 (s, 1H), 2.96-2.89 (m, 1H), 2.70 (d, J=4.7 Hz, 3H), 1.11 (d, J=6.9Hz, 6H). MS m/z (ESI): 167.1 [M+H]⁺.

Step 3:

C65-3 (1.14 g, 6.86 mmol) was dissolved in anhydrous DMF (10 mL), silveroxide (3.2 g, 13.7 mmol) and 2-bromoacetonitrile (905 mg, 7.55 mmol)were sequentially added to the solution, and the reaction was performedat room temperature for 4 h. The reaction solution was filtered, thefiltrate was diluted with ethyl acetate (30 mL), then washed with asaturated aqueous solution of sodium chloride (20 mL×3), dried overanhydrous sodium sulfate, and filtered. The filtrate was concentratedunder reduced pressure, and the residue was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=1:4), toafford C65-4 (837 mg, yellow solid, yield 59.4%).

¹H NMR (400 MHz, CDCl₃) δ 7.87 (s, 1H), 5.84 (s, 1H), 4.81 (s, 2H), 4.75(m, 1H), 3.08-3.02 (m, 1H), 2.92 (d, J=4.9 Hz, 3H), 1.20 (d, J=6.9 Hz,6H). MS m/z (ESI): 206.2 [M+H]⁺.

Step 4:

C65-4 (720 mg, 3.5 mmol), cyclopropylboronic acid (903 mg, 10.5 mmol),anhydrous cupric acetate (636 mg, 3.5 mmol) and DMAP (640 mg, 5.25 mmol)were added to acetonitrile (35 mL), and the reaction was placed in anoil bath at 50° C., and allowed to proceed for 18 h. The reactionsolution was filtered, the filtrate was concentrated, and then theresidue was purified by column chromatography on silica gel (petroleumether:ethyl acetate=4:1), to afford C65-5 (410 mg, colorless oil, yield47.6%).

¹H NMR (400 MHz, CDCl₃) δ 7.98 (s, 1H), 6.51 (s, 1H), 4.84 (s, 2H), 3.15(s, 3H), 3.10-3.02 (m, 1H), 2.54-2.49 (m, 1H), 1.21 (d, J=7.0 Hz, 6H),0.94-0.89 (m, 2H), 0.73-0.69 (m, 2H). MS m/z (ESI): 246.2 [M+H]⁺.

Step 5:

Compound C65-2 (800 mg, 4.4 mmol) was dissolved in DMF (5 mL), NaH (320mg, 13.2 mmol) was added at 0° C., and the reaction solution was stirredat 0° C. for 30 min. Then, iodoethane (687 mg, 4.4 mmol) was added, andthe reaction was stirred at room temperature for 18 hours. LC-MSindicated the reaction of the starting materials was complete. Thereaction was quenched by adding water (10 mL), and extracted with ethylacetate (20 mL×3). The organic phases were combined, washed withsaturated brine (40 mL), added with anhydrous sodium sulfate (10 g),dried for half an hour, and filtered. The filtrate was concentratedunder reduced pressure to afford a crude product, which was purified bycolumn chromatography on silica gel (petroleum ether:ethylacetate=20:1˜5:1) to afford compound C61-1 (360 mg, oil, yield 39.0%).MS m/z (ESI): 208.7 [M+H]⁺.

iii Preparation of Intermediate Example C119-1

Compound C22-2 (2.2 g, 0.010 mol) was dissolved in acetonitrile (30 mL),cyclopropylboronic acid (1.72 g, 0.020 mol), DMAP (1.8 g, 0.015 mol),molecular sieves (2.7 g) and cupric acetate (1.8 g, 0.010 mol) weresequentially added, and the reaction solution was stirred at 50° C. for48 hours. LC-MS indicated the reaction of the starting materials wassubstantially complete. The reaction solution was cooled to roomtemperature, filtered, and the filtrate was concentrated under reducedpressure to afford a crude product, which was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=10:1˜5:1) toafford compound C119-1 (1.2 g, yellow solid, yield 48%). MS m/z (ESI):306.8 [M+H]⁺.

iv Preparation of Intermediate Example C152-3

Step 1:

Compound C152-1 (5 g, 0.030 mol) was dissolved in dichloromethane (30mL), triethylamine (4.0 g, 0.040 mol) and trifluoroacetic anhydride (7.6g, 0.036 mol) were added, and the reaction solution was stirred at roomtemperature for 16 hours. The reaction solution was quenched by water(20 mL), and then extracted with dichloromethane (50 mL). The organicphase was washed with saturated brine (20 mL), dried, concentrated, andthe crude product was purified by column chromatography on silica gel(petroleum ether:ethyl acetate=10:1), to afford compound C152-2 (2.4 g,white solid, yield 30.5%).

¹H NMR (400 MHz, CDCl₃) δ 7.94 (s, 1H), 7.61 (s, 1H), 7.26 (s, 1H), 3.95(s, 3H), 3.24-3.13 (m, 1H), 1.24 (d, J=6.8 Hz, 6H). MS m/z (ESI): 262.9[M+H]⁺.

Step 2:

Compound C152-2 (2.4 g, 9.16 mmol) was dissolved in THE (30 mL), asolution of borane (27 mL, 27 mmol) was added, and the reaction solutionwas stirred at 50° C. overnight. The reaction was quenched by addingwater (5 mL), then extracted with ethyl acetate (20 mL×3). The organicphase was washed with water (15 mL), dried over sodium sulfate, and thenconcentrated to give a crude product, C152-3 (1.3 g, brown solid, yield57.2%). MS m/z (ESI): 248.9 [M+H]⁺.

Example 19: preparation of5-((5-bromo-2-methoxypyridin-4-yl)oxy)pyrimidine-2,4-diamine (C124,compound 124)

Step 1:

C124-1 (1.2 g, 9.6 mmol) was dissolved in acetic acid (50 mL), and thenan aqueous solution (10 mL) of KOH (2.15 g, 38.4 mmol) and bromine (3.07g, 19.2 mmol) were dropwise added. The reaction was performed at 0° C.for 0.5 hour, and was then stirred at room temperature for 16 hours. Alarge amount of white solid precipitated, the reaction was filtered, andthe filter cake was rinsed with petroleum ether (10 mL). The filter cakewas collected and dried to afford compound C124-2 (2.34 g, white solid,yield 86%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.14 (s, 1H), 3.88 (s, 3H). MS m/z (ESI):283.7 [M+H]⁺.

Step 2:

C124-2 (2.34 g, 8.26 mmol) was dissolved in tetrahydrofuran (50 mL), andthen a solution of n-butyl lithium (7.5 mL, 18.2 mmol) was dropwiseadded under the protection of nitrogen at −78° C., and the reaction wasperformed at −78° C. for 1 hour. The reaction was quenched by addingwater (10 mL), adjusted to pH 7 with a 1N solution of hydrochloric acid,and then extracted with ethyl acetate (40 mL×2). The organic phases werecombined, washed once with saturated brine (20 mL), then dried overanhydrous sodium sulfate (5 g) for half an hour, filtered, andconcentrated. The crude product was isolated by column chromatography onsilica gel (petroleum ether:ethyl acetate=3:1) to afford compound C124-3(1.2 g, white solid, yield 71.2%).

¹H NMR (400 MHz, DMSO-d₆) δ 11.53 (s, 1H), 8.09 (s, 1H), 3.78 (s, 3H).MS m/z (ESI): 203.8 [M+H]⁺.

Step 3:

C124-3 (204 mg, 1 mmol) was dissolved in DMF (2 mL), followed byaddition of potassium carbonate (276 mg, 2 mmol) and bromoacetonitrile(240 mg, 2 mmol), and the reaction was stirred at room temperature for 1h. The reaction was quenched by adding water (5 mL), and then extractedwith ethyl acetate (20 mL×2). The organic phase was dried over anhydroussodium sulfate, concentrated, and the crude product was isolated bycolumn chromatography on silica gel (petroleum ether:ethyl acetate=4:1)to afford compound C124-4 (100 mg, white solid, yield 43.3%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.19 (s, 1H), 6.32 (s, 1H), 4.85 (s, 3H),3.93 (s, 3H). MS m/z (ESI): 242.7 [M+H]⁺.

Step 4:

C124-4 (100 mg, 0.433 mmol) was dissolved in DMF (2 mL), and tert-butoxybis(dimethylamino)methane (226 mg, 1.3 mmol) was added to the reactionsolution. The reaction was stirred at 100° C. for 2 hours. After LC-MSindicated the reaction was complete, the reaction was quenched by addingwater, and extracted with ethyl acetate (15 mL×2). The organic phase wasdried over anhydrous sodium sulfate and concentrated to afford a crudeproduct, compound C124-5 (110 mg, crude). MS m/z (ESI): 297.8 [M−45+H]*.

Step 5:

C124-5 (242 mg, 0.705 mmol) was dissolved in DMF (2 mL), and anilinehydrobromide (24 mg, 0.14 mmol) was added to the reaction solution. Thereaction was stirred at 100° C. for 3 hours. The reaction was quenchedby adding water, and extracted with ethyl acetate (25 mL×3). The organicphase was dried and concentrated to afford compound C124-6 (110 mg,brown solid, yield 42.5%). MS m/z (ESI): 345.8 [M+H]⁺.

Step 6:

C124-6 (110 mg, 0.3 mmol) was dissolved in ethanol (5 mL), and guanidinehydrochloride (56 mg, 0.6 mmol) and sodium methoxide (32 mg, 0.6 mmol)were added to the reaction solution. The reaction was stirred at 90° C.overnight. After LC-MS indicated the reaction was complete, the reactionwas concentrated to dryness, and the residue was purified by thePrep-HPLC method to afford C124 (10 mg, yellow solid, yield 10%).

¹H NMR (400 MHz, CD₃OD) δ 8.23 (s, 1H), 7.23 (s, 1H), 6.37 (s, 1H), 3.88(s, 3H). MS m/z (ESI): 311.8 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 19.

Starting material or regent different Compound from that inCharacterization No. Compound Structure Name Example 19 Data C30(compound 30)

5-(2,5- dimethoxypyridin- 4-yl)oxy)pyrimidine- 2,4-diamine Thepreparation started from Step 3 of Example 19, and C124-3 in Step 3 wasreplaced with 2,5- dimethoxypyridin- 4-ol. ¹H NMR (400 MHz, CD₃OD) δ7.77 (s, 1H), 7.60 (s, 1H), 6.08 (s, 1H), 3.93 (s, 3H), 3.82 (s, 3H). MSm/z (ESI): 264.0 [M + H]⁺. C31 (compound 31)

5-((2-methoxy-4- yl)oxy)pyrimidine- 2,4-diamine The preparation startedfrom Step 3 of Example 19, and C124-3 in Step 3 was replaced with2-methoxypyridin- 4-ol. ¹H NMR (400 MHz, DMSO-d₆) δ 8.47 (s, 1H), 8.08(s, 2H), 7.97 (s, 1H), 7.69 (s, 2H), 6.78 (s, 1H), 6.47 (s, 1H), 3.84(s, 3H). MS m/z (ESI): 234.1 [M + H]⁺.

Example 20: preparation of(E)-(5-((2-methoxy-5-styrylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C183, compound 183)

C124 (32 mg, 0.1 mmol), (E)-styrylboronic acid (SM3, 29.6 mg, 0.2 mmol),potassium carbonate (41.4 mg, 0.3 mmol) and Pd(PPh₃)₄ (20 mg) were addedto a 50 mL single-neck flask, followed by addition of the solvent,1,4-dioxane (5 mL) and water (1 mL). Under the protection of nitrogen,the reaction solution was heated to 100° C., and the reaction wasperformed for 18 h. LC-MS analysis confirmed that the target product wasobtained. The reaction solution was cooled to room temperature,filtered, and the filtrate was concentrated under reduced pressure togive an oily crude product, which was purified by preparative thin layerchromatography on silica get (dichloromethane:methanol=10:1) to affordcompound C183 (6 mg, white solid, yield 17.9%).

¹H NMR (400 MHz, CD₃OD) δ 8.37 (s, 1H), 7.67 (s, 1H), 7.58 (d, J=8.0 Hz,2H), 7.35-7.38 (m, 3H), 7.25 (d, J=8.0 Hz, 2H), 6.05 (s, 1H), 3.90 (s,3H). MS m/z (ESI): 335.8 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 20.

Starting material or regent different Compound from that in No. CompoundStructure Name Example 20 Characterization Data C177 (compound 177)

4-((2,4- diamino- pyrimi din-5-yl) oxy)-6- methoxy- nicotinonitrilePotassium carbonate and SM3 were replaced with zinc powder and zinccyanide; 1,4-dioxane was replaced with DMF. ¹H NMR (400 MHz, CD₃OD) δ8.47 (s, 1H), 7.72 (s, 1H), 6.13 (s, 1H), 3.96 (s, 3H). MS m/z (ESI):259.0 [M + H]⁺. C176 (compound 176)

5-((2-methoxy- 5-methylpyridin- 4-yl)oxy) pyrimidine- 2,4-diamine SM3was replaced with methylboronic acid. ¹H NMR (400 MHz, CD₃OD) δ 7.87 (s,1H), 7.60 (s, 1H), 5.96 (s, 1H), 3.83 (s, 3H), 2.27 (s, 3H). MS m/z(ESI): 247.9 [M + H]⁺. C164 (compound 164)

5-((2-methoxy- 5-(prop-1-en-2- yl)pyridin-4- yl)oxy) pyrimidine-2,4-diamine SM3 was replaced with isopropenyl- boronic acid. ¹H NMR (400MHz, CD₃OD) δ 7.96 (s, 1H), 7.58 (s, 1H), 6.02 (s, 1H), 5.22 (d, J = 8.0Hz, 2H), 3.87 (s, 3H), 2.20 (s, 3H). MS m/z (ESI): 273.9 [M + H]⁺. C162(compound 162)

5-((2-methoxy- 5-vinylpyridin- 4-yl)oxy) pyrimidine- 2,4-diamine SM3 wasreplaced with 4,4,5,5- tetramethyl-2- vinyl-1,3,2- dioxaborolane. ¹H NMR(400 MHz, CD₃OD) δ 8.21 (s, 1H), 7.61 (s, 1H), 7.01- 6.92 (m, 1H), 6.01(s, 1H), 5.85 (d, J = 18.0 Hz, 1H), 5.33 (d, J = 11.6 Hz, 1H), 3.87 (s,3H). MS m/z (ESI): 259.9 [M + H]⁺. C163 (compound 163)

5-((2-methoxy- 5-phenylpyridin- 4-yl)oxy) pyrimidine- 2,4-diamine SM3was replaced with phenylboronic acid. ¹H NMR (400 MHz, CD₃OD) δ 8.07 (s,1H), 7.64-7.44 (m, 5H), 7.39-7.36 (m, 1H), 6.12 (s, 1H), 3.91 (s, 3H).MS m/z (ESI): 309.9 [M + H]⁺. C138 (compound 138)

(E)-5-((5- isopropyl-2- styrylpyridin-4- yl)oxy) pyrimidine- 2,4-diamineC124 was replaced with C2. ¹H NMR (400 MHz, DMSO) δ 8.40 (s, 1H),7.65-7.55 (m, 4H), 7.36-7.28 (m, 4H), 6.67 (s, 1H), 6.51 (s, 2H), 6.12(s, 2H), 3.34- 3.36 (m, 1H), 1.32 (d, J = 6.8 Hz, 6H). MS m/z (ESI):347.9 [M + H]⁺. C33 (compound 33)

5-((5- cyclopropyl-2- methoxypyridin- 4-yl)oxy) pyrimidine- 2,4-diamineSM3 was replaced with cyclopropyl- boronic acid. ¹H NMR (400 MHz, CD₃OD)δ 7.73 (s, 1H), 7.60 (s, 1H), 5.98 (s, 1H), 3.83 (s, 3H), 2.11- 2.08 (m,1H), 0.95- 0.74 ((m, 4H). MS m/z (ESI): 273.9 [M + H]⁺. C205 (compound205)

5-((5- (cyclopent-1- en-1-yl)-2- methoxypyridin- 4-yl)oxy) pyrimidine-2,4-diamine SM3 was replaced with cyclopentenyl- pinacol borate; thecatalyst and ligand was replaced with tri- tert- butylphosphine andPd₂(dba)₃; potassium carbonate was replaced with ¹H NMR (400 MHz, DMSO)δ 8.45 (s, 1H), 8.11 (s, 1H), 8.08 (s, 1H), 7.88 (s, 1H), 7.61 (s, 2H),6.38-6.35 (m, 2H), 3.83 (s, 3H), 2.74- 2.67 (m, 4H), 1.93- 1.89 (m, 2H).MS m/z (ESI): 299.9 [M + H]⁺. cesium carbonate; the solvent was dioxaneonly, no water was added. C54 (compound 54)

4-((2,4- diamino- pyrimidin-5- yl)oxy)- 5-isopropyl-2- cyanopyridinePotassium carbonate and SM3 were replaced with zinc powder and zinccyanide; 1,4-dioxane was replaced with DMF; C124 was replaced with C2.¹H NMR (400 MHz, CD₃OD) δ 8.52 (s, 1H), 7.63 (s, 1H), 7.09 (s, 1H),3.52-3.49 (m, 1H), 1.39 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 270.9 [M +H]⁺. C129 (compound 129)

4-((2,4- diamino- pyrimidin- 5-yl)oxy)-5- isopropyl- picolinamide It isa by-product isolated during the synthesis of C54. ¹H NMR (400 MHz,DMSO-d) δ 8.43 (s, 1H), 8.03 (s, 1H), 7.62 (s, 2H), 7.14 (s, 1H), 6.55(s, 2H), 6.15 (s, 2H), 3.43-3.41 (m, 1H), 1.32 (d, J = 6.8 Hz, 6H). MSm/z (ESI): 288.9 [M + H]⁺.

Example 21: preparation of1-(4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpyridin-2-yl)ethanone(C46, compound 46)

C45 (20 mg, 0.06 mmol) was dissolved in 2M HCl (10 mL), and the reactionwas performed at room temperature for 18 h. After the reaction wascomplete, the reaction solution was concentrated under reduced pressureto afford a crude product, which was purified by prep-HPLC to affordcompound C46 (5 mg, yield 27.47%).

1H NMR (400 MHz, CD₃OD) δ 8.60 (s, 1H), 7.75 (s, 1H), 7.38 (s, 1H),3.48-3.45 (m, 1H), 2.64 (s, 3H), 1.41 (d, J=6.0 Hz, 6H). MS m/z (ESI):287.9 [M+H]⁺.

Example 22: preparation of5-((5-isopropyl-2-((trimethylsilyl)ethynyl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C246, compound 246) and5-((2-ethynyl-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine (C66,compound 66)

Step 1:

Compound C2 (3 g, 9.29 mmol) was dissolved in 1,4-dioxane (40 mL),trimethylsilylacetylene (9 g, 92.9 mmol), DIEA (12 g, 92.9 mmol), CuI(0.6 g) and Pd(PPh₃)₂Cl₂ (0.6 g) were sequentially added, and purge withnitrogen was performed for 3 times. Under the protection of nitrogen,the reaction was performed at 50° C. for 2 hours. LC-MS indicated thereaction of the starting materials was substantially complete. Thereaction solution was cooled to room temperature, filtered, the filtercake was washed with 1,4-dioxane (10 mL), the filtrate was concentratedunder reduced pressure to remove dioxane, followed by addition ofpurified water (100 mL), and extraction with ethyl acetate (100 mL×3).The organic phases were combined, added with anhydrous sodium sulfate(20 g), dried for 30 min, filtered, and concentrated under reducedpressure to afford a crude product, which was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=20:1˜5:1) toafford compound C246 (2 g, yield 63.1%). MS m/z (ESI): 341.9 [M+H]⁺.

Step 2:

Compound C246 (2 g, 5.87 mmol) was dissolved in THE (20 mL), and TBAF(1.53 g, 5.87 mmol) was added. The reaction was performed at roomtemperature for 10 minutes. LC-MS indicated the reaction of the startingmaterials was complete. The reaction solution was rotary evaporated todryness to give an oily residue. The residue was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=1:3) toafford compound C66 (0.7 g, yellow solid, yield 44.6%).

¹H NMR (300 MHz, DMSO-d₆) δ 8.33 (s, 1H), 7.56 (s, 1H), 6.50 (s, 1H),6.41 (s, 2H), 6.01 (s, 2H), 4.20 (s, 1H), 3.37-3.31 (m, 1H), 1.28 (d,J=6.8 Hz, 6H). MS m/z (ESI): 269.8 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 22.

Starting material or regent different Compound from that in No. CompoundStructure Name Example 22 Characterization Data C88 (compound 88)

5-((2-ethynyl- 5-isopropyl- pyrdin-4- yl)thio) pyrimidine- 2,4-diamineC2 in Step 1 was replaced with C85. ¹H NMR (400 MHz, CD₃OD) δ 8.28 (s,1H), 7.96 (s, 1H), 6.80 (s, 1H), 3.66 (s, 1H), 3.37- 3.30 (m, 1H), 1.39(d, J = 6.8 Hz, 6H). MS m/z (ESI): 285.8 [M + H]⁺. C136 (compound 136)

5-((5- isopropyl-2- ((4- methoxyphenyl) ethynyl)pyridin- 4-yl)oxy)pyrimidine- 2,4-diamine Trimethylsilylace tylene in Step 1 was replacedwith 4- methoxy- phenylacetylene. ¹H NMR (400 MHz, DMSO-d₆) δ 8.50- 8.46(m, 2H), 8.24 (s, 1H), 8.10 (s, 1H), 7.95 (s, 2H), 7.52 (d, J = 8.8 Hz,2H), 7.19 (s, 1H), 7.01 (d, J = 8.8 Hz, 2H), 3.81 (s, 3H), 3.35 (m, 1H),1.30 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 375.9 [M + H]⁺. C137 (compound137)

4-((4-(2,4- diaminopyrimidin- 5-yl)oxy)-5- isopropylpyridin-2-yl)ethynyl) benzonitrile Trimethylsilylace tylene in Step 1 wasreplaced with 4- ethynyl- benzonitrile. ¹H NMR (400 MHz, DMSO-d₆) δ 8.51(s, 2H), 8.11 (s, 1H), 7.98 (s, 1H), 7.92 (d, J = 8.4 Hz, 2H), 7.78 (d,J = 8.8 Hz, 2H), 7.68 (s, 2H), 7.31 (s, 1H), 3.38 (m, 1H), 1.32 (d, J =6.8 Hz, 6H). MS m/z (ESI): 370.9 [M + H]⁺. C148 (compound 148)

5-((2-((4- (dimethylamino) phenyl) ethynyl)-5- isopropylpyridin-4-yl)oxy) pyrimidine-2,4- diamine Trimethylsilylace tylene in Step 1 wasreplaced with 4- dimethylamino- phenylacetylene. ¹H NMR (400 MHz,CDCl₃)) δ 8.42 (s, 1H), 7.76 (s, 1H), 7.47 (d, J = 8.4 Hz, 2H), 6.80 (s,1H), 6.65 (d, J = 8.8 Hz, 2H), 5.09 (s, 2H), 4.92 (s, 2H), 3.38 (m, 1H),3.02 (s, 6H), 1.39 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 388.9 [M + H]⁺.C155 (compound 155)

2-(4-(2,4- diamino- pyrimidin- 5-yl)oxy)-5- isopropylpyridin-2-yl)-1-(4- (dimethylamino) phenyl)ethanone It is a by-product resultedfrom the work-up of C148. ¹H NMR (400 MHz, DMSO-d₆) δ 8.26 (s, 1H), 7.83(m, 2H), 7.52 (s, 1H), 6.70 (m, 2H), 6.54 (s, 1H), 6.36 (s, 2H), 5.96(s, 2H), 4.22 (s, 2H), 3.32 (m, 1H), 2.95 (s, 6H), 1.27 (d, J = 6.8 Hz,6H). MS m/z (ESI): 406.7 [M + H]⁺. C170 (compound 170)

5-((2- methoxy-5- ((trimethylsilyl) ethynyl) pyridin-4- yl)oxy)pyrimidine- 2,4-diamine C2 in Step 1 was replaced with C124. ¹H NMR (400MHz, CDCl₃) δ 8.24 (s, 1H), 7.65 (s, 1H), 6.15 (s, 1H), 5.73 (s, 2H),5.28 (s, 2H), 3.92 (s, 3H), 0.25 (s, 9H). MS m/z (ESI): 330.0 [M + H]⁺.C126 (compound 126)

4-(4-(2,4- diamino- pyrimidin-5- yl)oxy)-5- isopropyl- pyridin- 2-yl)-2-methylbut-3- yn-2-ol Trimethylsilylace tylene in Step 1 was replacedwith 2- methylbut-3-yn- 2-ol; 1,4-dioxane was replaced withacetonitrile; DIEA was replaced with triethylamine. ¹H NMR (400 MHz,DMSO-d₆) δ 8.37 (s, 1H), 6.54-6.48 (m, 3H), 6.09 (s, 2H), 5.54 (s, 1H),3.39-3.33 (m, 1H), 1.42 (s, 6H), 1.30 (d, J = 7.2 Hz, 6H). MS m/z (ESI):327.9 [M + H]⁺. C171 (compound 171)

5-((5-ethynyl- 2-methoxy- pyridin-4- yl)oxy) pyrimidine- 2,4-diamine Thepreparation started from Step 2, C246 in Step 2 was replaced with C170.¹H NMR (400 MHz, CD₃OD) δ 8.47 (s, 1H), 7.64 (s, 1H), 6.06 (s, 1H), 3.89(s, 3H), 3.68 (s, 1H). MS m/z (ESI): 257.9 [M + H]⁺. C143 (compound 143)

3-(4-((2,4- diaminopyrim idin-5- yl)oxy)-5- isopropylpyri din-2-yl)prop-2-yn-1-ol Trimethylsilylace tylene in Step 1 was replaced withprop-2-yn- 1-ol. ¹H NMR (400 MHz, DMSO-d₆) δ 8.36 (s, 1H), 7.59 (s, 1H),6.48 (s, 1H), 6.45 (s, 2H), 6.05 (s, 2H), 5.35- 5.32 (m, 1H), 4.26- 4.25(m, 2H), 3.38- 3.33 (m, 1H), 1.30 (d, J = 6.8 Hz, 6H). MS m/z (ESI):299.9 [M + H]⁺. C142 (compound 142)

5-((2- (cyclo- propylethynyl)-5- isopropylpyrdin- 4-yl)oxy)pyrimidine-2,4- diamine Trimethylsilylace tylene in Step 1 was replacedwith cyclo- propylacetylene. ¹H NMR (400 MHz, DMSO-d₆) δ 8.46- 8.37 (m,2H), 8.04 (s, 1H), 7.90 (s, 1H), 7.63 (s, 2H), 7.00 (s, 1H), 3.33-3.29(m, 1H), 1.55-1.52 (m, 1H), 1.27 (d, J = 6.8 Hz, 6H), 0.91-0.89 (m, 2H),0.76-0.74 (m, 2H). MS m/z (ESI): 310.1 [M + H]⁺. C215 (compound 215)

5-((2-ethynyl- 5-(prop-1-en- 2-yl)pyridin- 4-yl)oxy) pyrimidine-2,4-diamine C2 in Step 1 was replaced with C215-1. ¹H NMR (400 MHz, DMSO-d₆)δ 8.48 (bs, 1H), 8.42 (s, 1H), 8.02 (s, 1H), 7.92 (s, 1H), 7.69 (bs,2H), 7.67 (s, 1H), 5.35 (d, J = 7.2 Hz, 2H), 4.38 (s, 1H), 2.15 (s, 3H).MS m/z (ESI): 267.8 [M + H]⁺. C134 (compound 134)

5-((5- isopropyl-2- (prop-1-yn-1- yl)pyridin-4- yl)oxy) pyrimidine-2,4-diamine Trimethylsilylace tylene in Step 1 was replaced with propyne. ¹HNMR (400 MHz, DMSO-d₆) δ 8.31 (s, 1H), 7.60 (s, 1H), 6.58 (s, 2H), 6.48(s, 1H), 6.17 (s, 2H), 3.36- 3.31 (m, 1H), 2.00 (s, 3H), 1.28 (d, J =6.8 Hz, 6H). MS m/z (ESI): 283.8 [M + H]⁺. C135 (compound 135)

5-((5- isopropyl-2- (phenyl- ethynyl)pyridin- 4-yl)oxy) pyrimidine-2,4-diamine Trimethylsilylace tylene in Step 1 was replaced withphenylacetylene. ¹H NMR (400 MHz, DMSO-d₆) δ 8.46 (s, 2H), 8.08 (s, 1H),7.95 (s, 1H), 6.65-7.55 (m, 4H), 7.73-7.42 (m, 3H) 7.25 (s, 1H), 3.42-3.26 (m, 1H), 1.31 (d, J = 8.8 Hz, 6H). MS m/z (ESI): 346.2 [M + H]⁺.C228 (compound 228)

2-((4-amino- 5-((2-ethynyl- 5-isopropyl- pyrdin-4- yl)oxy) pyrimidin-2-yl)amino) propane-1,3-diol C2 in Step 1 was replaced with C111. ¹H NMR(400 MHz, CD₃OD) δ 8.34 (s, 1H), 7.65 (s, 1H), 6.77 (s, 1H), 3.74-3.73(m, 1H), 3.70 (d, 4H), 3.50- 3.49 (m, 1H), 3.47- 3.46 (m, 1H), 1.40 (d,J = 6.8 Hz, 6H). MS m/z (ESI): 343.8 [M + H]⁺. C231 (compound 231)

5-((2-ethynyl- 5-isopropyl- pyridin-4-yl)oxy)- N²-methyl-pyrimidine-2,4- diamine C2 in Step 1 was replaced with C237. ¹H NMR (400MHz, CD₃OD) δ 8.33 (s, 1H), 7.63 (s, 1H), 6.76 (s, 1H), 3.69 (s, 1H),3.45- 3.40 (m, 1H), 2.91 (s, 3H), 1.40 (d, J = 5.2 Hz, 6H). MS m/z(ESI): 283.9 [M + H]⁺. C230 (compound 230)

2-((4-amino- 5-((2-ethynyl- 5-isopropyl- pyridin-4- yl)oxy) pyrimidin-2-yl)amino) ethanol C2 in Step 1 was replaced with C236. ¹H NMR (400 MHz,CD₃OD) δ 8.33 (s, 1H), 7.64 (s, 1H), 6.76 (s, 1H), 3.74-3.72 (m, 3H),3.49-3.47 (m, 3H), 1.39 (m, J = 6.8 Hz, 6H). MS m/z (ESI): 313.9 [M +H]⁺. C229 (compound 229)

3-((4-amino- 5-((2-ethynyl- 5-isopropyl- pyridin-4- yl)oxy) pyrimidin-2-yl)amino) propane-l,2-diol C2 in Step 1 was replaced with C235. ¹H NMR(400 MHz, CD₃OD) δ 8.33 (s, 1H), 7.64 (s, 1H), 6.75 (s, 1H), 3.81-3.79(m, 1H), 3.70 (s, 1H), 3.46- 3.41 (m, 2H), 3.39- 3.34 (m, 3H), 1.40 (d,J = 6.8 Hz, 6H). MS m/z (ESI): 343.8 [M + H]⁺. C233 (compound 233)

5-((2-ethynyl- 5-isopropyl- pyrdin-4-yl)oxy)- N²-isopropyl-pyrmidine-2,4- diamine C2 in Step 1 was replaced with C239. ¹H NMR (400MHz, CD₃OD) δ 8.33 (s, 1H), 7.62 (s, 1H), 6.77 (s, 1H), 4.09-4.06 (m,1H), 3.69 (s, 1H), 3.47 (m, 1H), 1.41-1.38 (m, 6H), 1.24-1.21 (m, 6H).MS m/z (ESI): 311.9 [M + H]⁺.

Example 23: preparation of5-((2-bromo-5-isopropylpyridin-4-yl)oxy)-N⁴-(2-(ethyl(methyl)amino)ethyl)pyrimidine-2,4-diamine(C154, compound 154)

Step 1:

Compound C2-5 (5 g, 0.023 mol) was dissolved in acetone (200 mL), ethylbromoacetate (5.8 g, 0.035 mol) and potassium carbonate (6.4 g, 0.046mol) were sequentially added, and the reaction solution was stirred at50° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was quenched by adding water (100mL), and extracted with ethyl acetate (100 mL×3). The organic phaseswere combined, washed once with saturated brine (100 mL), then driedover anhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated, to afford a crude product, compound C154-2, which wasdirectly used in the next step (5 g crude product, brown oily liquid,yield 74%). MS m/z (ESI): 288.0 [M+H]⁺.

Step 2:

Compound C154-2 (4.5 g, 0.015 mol) was dissolved in DMF (30 mL), DMF-DMA(5.6 g, 0.047 mol) was added, and the reaction solution was stirred at130° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was quenched by adding water (100mL), and extracted with ethyl acetate (100 mL×3). The organic phaseswere combined, washed once with saturated brine (100 mL), then driedover anhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=10:1˜5:1) to afford compound C154-3 (2 g, yellow oily liquid,yield 40%). MS m/z (ESI): 342.8 [M+H]⁺.

Step 3:

Compound C154-3 (2 g, 0.0056 mol) was dissolved in ethanol (20 mL),guanidine hydrochloride (3.2 g, 0.034 mol) and sodium methoxide (1.82 g,0.034 mol) were sequentially added, and the reaction solution wasstirred at 90° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was filtered, thefiltrate was concentrated under reduced pressure, added with purifiedwater (50 mL), then adjusted to a pH value of 7 with 1 M HCl, andextracted with ethyl acetate (100 mL×3). The organic phases werecombined, washed once with saturated brine (100 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated to afford a crude product, which was isolated by columnchromatography on silica gel (petroleum ether:ethyl acetate=20:1˜5:1) toafford compound C154-4 (0.7 g, light yellow solid, yield 38%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.19 (s, 1H), 7.88 (s, 1H), 7.67 (s, 2H),7.16 (s, 1H), 3.25-3.18 (m, 1H), 1.27 (d, J=6.8 Hz, 6H). MS m/z (ESI):324.8 [M+H]⁺.

Step 4:

Compound C154-4 (200 mg, 0.617 mmol) was dissolved in phosphorusoxychloride (1 mL), heated to 80° C., and stirred for 16 hours. LC-MSindicated the reaction of the starting materials was complete. Thereaction solution was cooled to room temperature, quenched by addingwater (30 mL), then adjusted to a pH value of 7-8 with sodiumbicarbonate, and extracted with dichloromethane (50 mL×3). The organicphases were combined, washed with saturated brine (50 mL), added withanhydrous sodium sulfate, dried for half an hour, and filtered. Thefiltrate was concentrated under reduced pressure, and the crude productwas isolated by column chromatography on silica gel (petroleumether:ethyl acetate=10:1˜1:1) to afford compound C154-5 (90 mg, lightyellow solid, yield 42%).

¹H NMR (400 MHz, CDCl₃) δ 8.24 (s, 1H), 8.16 (s, 1H), 6.58 (s, 1H), 5.35(s, 2H), 3.38-3.35 (m, 1H), 1.38 (d, J=6.8 Hz, 6H). MS m/z (ESI): 342.7[M+H]⁺.

Step 5:

Compound C154-5 (10 mg, 0.029 mmol) was dissolved in acetonitrile (2mL), N-methyl-N-ethylethylenediamine (12.4 mg, 0.18 mmol) was added, andthe reaction solution was stirred at 90° C. for 16 hours. LC-MSindicated the reaction of the starting materials was substantiallycomplete. The reaction solution was concentrated under reduced pressureto afford a crude product, which was purified by preparative thin layerchromatography on silica gel (dichloromethane:methanol=10:1) to affordcompound C154 (3 mg, white solid, yield 31%).

¹H NMR (400 MHz, CD₃OD) δ 8.18 (s, 1H), 7.59 (s, 1H), 6.77 (s, 1H), 3.61(m, 2H), 3.34-3.32 (m, 1H), 2.85-2.77 (m, 4H), 2.50 (s, 3H), 1.38 (d,J=6.8 Hz, 6H), 1.14 (s, 3H). MS m/z (ESI): 410.8 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 23.

Starting material or regent different Compound Compound from that in No.Structure Name Example 23 Characterization Data C71 (compound 71)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴-(2- (dimethylamino)ethyl)pyrimidine- 2,4-diamine N-methyl-N- ethylethylene diamine in Step5 was replaced with N,N- dimethylethyl enediamine. ¹H NMR (400 MHz,CD₃OD) δ 8.19 (s, 1H), 7.62 (s, 1H), 6.82 (s, 1H), 3.74-3.71 (m, 2H),3.37-3.34 (m, 1H), 3.12 (s, 2H), 2.78 (s, 6H), 1.38 (d, J = 6.8 Hz, 6H).MS m/z (ESI): 394.9 [M + H]⁺. C153 (compound 153)

2-((2-amino-5- ((2-bromo-5- isopropylpyridin- 4-yl)oxy) pyrimidin-4-yl)amino) propane-1,3-diol N-methyl-N- ethylethylene diamine in Step 5was replaced with serinol. ¹H NMR (400 MHz, CD₃OD) δ 8.18 (s, 1H), 7.61(s, 1H), 6.89 (s, 1H), 4.30 (s, 1H), 3.68- 3.60 (m, 4H), 3.31 (s, 1H),1.36 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 397.8 [M + H]⁺. C172 (compound172)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴- methyl- pyrimidine-2,4-diamine N-methyl-N- ethylethylene diamine in Step 5 was replacedwith methylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.17 (s, 1H), 7.52 (s, 1H),6.73 (s, 1H), 3.41 (s, 1H), 2.92 (s, 3H), 1.37 (d, J = 6.8 Hz, 6H). MSm/z (ESI): 337.8 [M + H]⁺. C173 (compound 173)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴- ethylpyrimidine-2,4-diamine N-methyl-N- ethylethylene diamine in Step 5 was replacedwith ethylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.17 (s, 1H), 7.52 (s, 1H),6.73 (s, 1H), 3.50-3.45 (m, 3H), 1.37 (m, J = 6.8 Hz, 6H), 1.19-1.12 (m,3H). MS m/z (ESI): 351.8 [M + H]⁺. C174 (compound 174)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴- isopropyl- pyrimidine-2,4-diamine N-methyl-N- ethylethylene diamine in Step 5 was replacedwith isopropylamine. ¹H NMR (400 MHz, CDCl₃) δ 8.28 (s, 1H), 7.54 (s,1H), 6.83 (s, 1H), 4.33-4.30 (m, 1H), 3.31-3.27 (m, 1H), 1.36 (d, J =6.8 Hz, 6H), 1.26 (d, J = 6.4 Hz, 6H). MS m/z (ESI): 365.8 [M + H]⁺.C175 (compound 175)

2-((2-amino-5- ((2-bromo-5- isopropylpyridin- 4-yl)oxy) pyrimidin-4-yl)amino)ethanol. N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with ethanolamine. ¹H NMR (400 MHz, CD₃OD) δ 8.17 (s, 1H), 7.56(s, 1H), 6.79 (s, 1H), 3.67-3.69 (m, 2H), 3.57-3.54 (m, 2H), 3.44- 3.37(m, 1H), 1.38 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 367.8 [M + H]⁺. C179(compound 179)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴- cyclopropyl-pyrimidine-2,4- diamine N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with cyclopropylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.16 (s, 1H),7.53 (s, 1H), 6.71 (s, 1H), 3.42-3.39 (m, 1H), 2.82-2.80 (m, 1H), 1.37-1.31 (d, J = 7.2 Hz, 6H), 0.79- 0.76 (m, 2H), 0.55 (s, 2H). MS m/z(ESI): 363.7 [M + H]⁺. C180 (compound 180)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴- cyclohexyl-pyrimidine-2,4- diamine N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with cyclohexylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.17 (s, 1H),7.52 (s, 1H), 6.74 (s, 1H), 4.07-4.01 (m, 1H), 3.46-3.39 (m, 1H), 1.92(d, J = 12.4 Hz, 2H), 1.78 (d, J = 13.2 Hz, 2H), 1.67 (d, J = 13.2 Hz,2H), 1.38-1.36 (m, 10H). MS m/z (ESI): 407.8 [M + H]⁺. C181 (compound181)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴- isobutyl- pyrimidine-2,4-diamine N-methyl-N- ethylethylene diamine in Step 5 was replacedwith isobutylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.19 (s, 1H), 7.53 (s,1H), 6.75 (s, 1H), 3.44-3.39 (m, 1H), 3.24 (d, J = 6.8 Hz, 2H),1.96-1.93 (m, 1H), 1.38 (d, J = 7.2 Hz, 6H), 0.90 (d, J = 6.8 Hz, 6H).MS m/z (ESI): 381.8 [M + H]⁺. C182 (compound 182)

(R)-5-((2-bromo- 5-isopropyl pyridin-4-yl)oxy)- N⁴-(1-cyclopro-pylethyl) pyrimidine-2,4- diamine N-methyl-N- ethylethylene diamine inStep 5 was replaced with (R)-1-cyclopro- pylethylamine. ¹H NMR (300 MHz,CD₃OD) δ 8.13 (s, 1H), 7.48 (s, 1H), 6.70 (s, 1H), 3.65 (m, 1H),3.42-3.40 (m, 1H), 1.36 (d, J = 4.5 Hz, 6H), 1.29 (s, 3H), 0.92-0.90 (m,1H), 0.49- 0.43 (m, 2H), 0.32-0.21 (m, 2H). MS m/z (ESI): 393.9 [M +H]⁺. C201 (compound 201)

2-(2-((2-amino- 5-((2-bromo-5- isopropylpyridin- 4-yl)oxy) pyrimidin-4-yl)amino)ethoxy) ethanol N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with 2-(2- aminoethoxy) ethanol. ¹H NMR (400 MHz, CD₃OD) δ 8.18(s, 1H), 7.56 (s, 1H), 6.76 (s, 1H), 3.63-3.61 (m, 6H), 3.55-3.54 (m,2H), 3.44- 3.42 (m, 1H), 1.38 (d, J = 7.2 Hz, 6H). MS m/z (ESI): 411.7[M + H]⁺. C202 (compound 202)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴-(2- (methylamino)ethyl)pyrimidine- 2,4-diamine N-methyl-N- ethylethylene diamine in Step5 was replaced with N-methyl- ethylenediamine. ¹H NMR (400 MHz, CD₃OD) δ8.28 (s, 1H), 7.83 (s, 1H), 7.23 (s, 1H), 3.89-3.86 (m, 2H), 3.37-3.35(m, 1H), 3.35- 3.33 (m, 2H), 2.81 (s, 3H), 1.37 (d, J = 7.2 Hz, 6H). MSm/z (ESI): 380.8 [M + H]⁺. C203 (compound 203)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴-(2- morpholinoethyl)pyrimidine-2,4- diamine N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with 2-morpholino- ethylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.20(s, 1H), 7.58 (s, 1H), 6.77 (s, 1H), 3.60 (m, 6H), 3.44-3.41 (m, 1H),2.57- 2.49 (m, 6H), 1.40 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 436.7 [M +H]⁺. C204 (compound 204)

3-((2-amino-5- ((2-bromo-5- isopropylpyridin- 4-yl)oxy) pyrimidin-4-yl)amino)propane- 1,2-diol N-methyl-N- ethylethylene diamine in Step 5was replaced with 3-amino propane-1,2-diol. ¹H NMR (400 MHz, CD₃OD) δ8.26 (s, 1H), 7.75 (s, 1H), 7.19 (s, 1H), 3.89 (m, 1H), 3.74-3.71 (m,1H), 3.61- 3.56 (m, 3H), 3.41-3.38 (m, 1H), 1.37 (d, J = 7.2 Hz, 6H). MSm/z (ESI): 399.7 [M + H]⁺. C207 (compound 207)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴-(2- methoxyethyl)pyrimidine-2,4- diamine N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with 2-methoxyethyl amine. ¹H NMR (400 MHz, CD₃OD) δ 8.18 (s,1H), 7.56 (s, 1H), 6.76 (s, 1H), 3.62-3.60 (m, 2H), 3.54-3.52 (m, 2H),3.42 (m, 1H), 3.35-3.33 (m, 3H), 1.38 (d, J = 6.8 Hz, 6H). MS m/z (ESI):383.7 [M + H]⁺. C209 (compound 209)

(S)-2-((2-amino- 5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)pyrimidin-4-yl) amino)but-1-ol N-methyl-N- ethylethylene diamine in Step5 was replaced with (S)-2- aminobut-1-ol. ¹H NMR (400 MHz, CD₃OD) δ 8.18(s, 1H), 7.57 (s, 1H), 6.82 (s, 1H), 3.60-3.58 (m, 2H), 3.45-3.40 (m,1H), 1.53- 1.47 (m, 1H), 1.40-1.38 (m, 2H), 1.37-1.31 (m, 6H), 0.95-0.92 (m, 3H). MS m/z (ESI): 395.8 [M + H]⁺. C210 (compound 210)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N⁴-(2- (pyrrolidin-1-yl)ethyl) pyrimidine-2,4- diamine N-methyl-N- ethylethylene diamine inStep 5 was replaced with 1-(2-aminoethyl) pyrrolidine. ¹H NMR (400 MHz,CD₃OD) δ 8.19 (s, 1H), 7.59 (s, 1H), 6.77 (s, 1H), 3.64 (s, 2H), 3.50(s, 1H), 2.92-2.78 (s, 6H), 1.89 (s, 4H), 1.38 (d, J = 6.4 Hz, 6H). MSm/z (ESI): 423.1 [M + H]⁺. C222 (compound 222)

tert-butyl 2-(2- amino-5-((2- bromo-5- isopropylpyridin- 4-yl)oxy)pyrimidin- 4-yl)hydrazine- 1-carboxylate N-methyl-N- ethylethylenediamine in Step 5 was replaced with tert-butyl hydrazine- carboxylate.¹H NMR (400 MHz, CD₃OD) δ 8.17 (s, 1H), 7.66 (s, 1H), 6.92 (s, 1H),3.43-3.41 (s, 1H), 1.50 (s, 9H), 1.37 (d, J = 6.8 Hz, 6H). MS m/z (ESI):438.9 [M + H]⁺. C241 (compound 241)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-4-(2- phenylhydrazineyl)pyrimidin-2- amine N-methyl-N- ethylethylene diamine in Step 5 wasreplaced with phenylhydrazine. ¹H NMR (400 MHz, CD₃OD) δ 7.99 (s, 1H),7.46 (s, 5H), 7.35-7.32 (m, 1H), 7.14- 7.12 (m, 1H), 3.10-3.05 (m, 1H),1.35 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 414.7 [M + H]⁺.

Example 24: preparation of5-((2-bromo-5-isopropylpyridin-4-yl)thio)pyrimidine-2,4-diamine (C85,compound 85)

Step 1:

Compound C85-1 (100 g, 0.58 mol) was dissolved in 1,4-dioxane (700 mL),isopropenylpinacol borate (147 g, 0.87 mol), K₂CO₃ (160 g, 1.16 mol) andPd(PPh₃)₄ (10 g, 5.4 mmol) were sequentially added, followed by additionof purified water (35 mL). Under the protection of nitrogen, thereaction was performed at 100° C. for 18 hours. LC-MS indicated thereaction of the starting materials was substantially complete, and thetarget product was obtained. The reaction solution was cooled to roomtemperature, filtered, the filter cake was washed with 1,4-dioxane (200mL), concentrated under reduced pressure to remove the organic solvent,added with purified water (200 mL), and extracted with ethyl acetate(400 ml×3). The organic phases were combined, added with 100 g anhydroussodium sulfate, dried for 30 min, and filtered. The filter cake waswashed with 200 ml ethyl acetate, concentrated under reduced pressure todryness, and the crude product was purified by column chromatography onsilica gel (mobile phase: petroleum ether:ethyl acetate=20:1˜2:1), toafford compound C85-2 (70 g, light yellow solid, yield 89.60%). MS m/z(ESI): 135.1 [M+H]⁺.

Step 2:

Compound C2-2 (70 g, 0.52 mol) was dissolved in anhydrous methanol (700mL), 10% palladium/carbon (14 g) was added, and the reaction wasperformed under hydrogen (0.4 MPa) at room temperature for 18 hours.LC-MS indicated a small amount of the starting material remained.Palladium/carbon 4 g was supplemented, and the reaction was continuedunder hydrogen (0.4 MPa) at room temperature for 18 hours. LC-MSindicated the reaction of the starting materials was complete. Thereaction solution was filtered, the filter cake was washed with 100 mlmethanol, and concentrated under reduced pressure to give a crudeproduct, compound C85-3 (70 g, orange oily liquid, yield 98.1%). MS m/z(ESI): 137.1 [M+H]⁺.

Step 3:

C85-3 (70 g, 0.52 mol) was dissolved in hydrobromic acid (40%) (400 mL).Under the protection of nitrogen, the reaction was cooled to −15°C.˜−20° C., and liquid bromine (200 g, 1.25 mol) was slowly dropwiseadded. After the dropwise addition was complete, the reaction was keptat the temperature for 1 h, sodium nitrite (103.5 g, 1.5 mol) wasdissolved in water (90 mL), and this solution was slowly dropwise added.After the dropwise addition was complete, the reaction was stirred atroom temperature for 18 h. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was adjusted to pH 8˜10 with asaturated solution of sodium bicarbonate, and extracted with ethylacetate (200 ml×3). The organic phases were combined, washed once withsaturated brine (200 mL), then dried over anhydrous sodium sulfate (400g) for half an hour, filtered, and concentrated. The crude product waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=50:1˜20:1), to afford compound C85-4 (30 g, light yellow oilyliquid, yield 29.4%). MS m/z (ESI): 199.8 [M+H]⁺.

Step 4:

Compound C85-4 (30 g, 0.15 mol) was dissolved in dichloromethane (300mL). Under the protection of nitrogen, meta-chloroperbenzoic acid (52 g,0.3 mol) was added at room temperature. After the addition, the reactionwas performed for 18 hours. LC-MS indicated the reaction of the startingmaterials was complete (if there is remaining starting material,meta-chloroperbenzoic acid can supplemented appropriately). The reactionwas filtered, the filter cake was washed twice with dichloromethane (100mL), the filtrate was concentrated under reduced pressure to dryness toafford compound C85-5 (30 g, light-colored solid, yield 92.6%).

¹H NMR (400 MHz, DMSO) δ 8.41 (s, 1H), 7.82 (d, J=6.8 Hz, 1H), 7.19 (d,J=8.4 Hz, 1H), 4.07-4.06 (m, 1H), 1.19 (d, J=6.8 Hz, 6H). MS m/z (ESI):215.9 [M+H]⁺.

Step 5:

Compound C85-5 (30 g, 0.14 mol) was slowly added to concentratedsulfuric acid (50 mL), the reaction was cooled to 0° C., fuming nitricacid (100 mL) was slowly dropwise added, and the reaction was heated to100° C. and allowed to proceed for 18 h. LC-MS indicated the reaction ofthe starting materials was complete. The reaction solution was cooled toroom temperature, slowly poured into ice water, adjusted to pH 9 with asolution of sodium hydroxide, then extracted with ethyl acetate (300ml×3). The organic phases were combined, dried over sodium sulfate, andthen filtered. The filtrate was concentrated under reduced pressure toafford a crude product, which was then purified by column chromatographyon silica gel (petroleum ether:ethyl acetate=20:1˜5:1) to afford theproduct C85-6 (25 g, oily liquid, yield 68.5%). MS m/z (ESI): 260.8[M+H]⁺.

Step 6:

Compound C85-6 (20 g, 0.08 mol) was dissolved in tetrahydrofuran (50mL), methanol (50 mL) and water (50 mL), iron powder (22.4 g, 0.4 mol)and ammonium chloride (21.6 g, 0.4 mol) were added at room temperature,and the reaction solution was heated to reflux for 18 h. LC-MS indicatedthe reaction of the starting materials was complete. The reactionsolution was cooled to room temperature, filtered, and the filtrate wasconcentrated under reduced pressure to dryness to afford compound C85-7(12 g, oily liquid, yield 62.8%). MS m/z (ESI): 214.9 [M+H]⁺.

Step 7:

Compound C85-7 (10 g, 0.047 mol) was dissolved in acetonitrile (100 mL),followed by addition of ethyl mercaptoacetate (9.9 g, 0.094 mol), thereaction was heated to 50° C., and stirred for 0.5 hour. Tert-butylnitrite was slowly added, and after completion of the addition, thereaction was stirred at the former temperature for 18 h. LC-MS indicatedthe reaction of the starting materials was complete. The reaction wascooled to room temperature, added with 50 ml water, and extracted withethyl acetate (100 ml×3). The organic phase was washed with saturatedbrine (20 mL×3), added with anhydrous sodium sulfate, dried for half anhour, and filtered. The filtrate was concentrated under reducedpressure, and the crude product was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=20:1˜5:1), to afford compoundC85-8 (8 g, oily liquid, yield 56.8%). MS m/z (ESI): 303.8 [M+H]⁺.

Step 8:

Compound C85-8 (8 g, 0.026 mol) was dissolved in anhydrous methanol (100mL). Under the protection of nitrogen, a solution of ammonia in methanol(7 mol/L, 40 mL) was added, and the reaction was stirred at roomtemperature for 18 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was concentrated under reducedpressure to afford compound C85-9 (6.5 g, light yellow solid, yield86.7%). MS m/z (ESI): 288.8 [M+H]⁺.

Step 9:

Compound C85-9 (6.5 g, 0.023 mol) was dissolved in anhydrousdichloromethane (50 mL), triethylamine (9.3 g, 0.092 mol) was added, thereaction solution was cooled to 0° C., trifluoroacetic anhydride (9.7 g,0.046 mol) was slowly dropwise added, and the reaction was stirred atroom temperature for 1 hour. LC-MS indicated the reaction of thestarting materials was complete. The reaction was added with water (50mL), extracted with dichloromethane (50 mL×3), dried over sodiumsulfate, and then filtered. The filtrate was concentrated under reducedpressure, and the crude product was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=20:1˜10:1) to afford compoundC85-10 (3.5 g, light yellow solid, yield 56.4%). MS m/z (ESI): 270.8[M+H]⁺.

Step 10:

Compound C85-10 (3.5 g, 0.013 mol) was dissolved in anhydrous DMF (30mL), tert-butoxy bis(dimethylamino)methane (6.8 g, 0.039 mol) was added,and the reaction solution was heated to 100° C., and stirred for 2hours. LC-MS indicated the reaction of the starting materials wascomplete. The reaction was cooled to room temperature, added with water(100 mL), extracted with ethyl acetate (50 mL×3), dried over sodiumsulfate, and then filtered. The filtrate was concentrated under reducedpressure, and the crude product was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=20:1˜5:1) to afford compoundC85-11 (2.2 g, light yellow oily liquid, yield 45.7%). MS m/z (ESI):325.8 [M+H]⁺.

Step 11:

Compound C85-11 (2.2 g, 0.006 mol) was dissolved in anhydrous DMF (20mL), aniline hydrobromide (1.6 g, 0.009 mol) was added, and the reactionsolution was heated to 100° C., and stirred for 18 hours. LC-MSindicated the reaction of the starting materials was complete. Thereaction was cooled to room temperature, added with water (20 mL),extracted with ethyl acetate (20 mL×3), washed twice with saturatedbrine, dried over sodium sulfate, and then filtered. The filtrate wasconcentrated under reduced pressure, and the crude product was purifiedby column chromatography on silica gel (petroleum ether:ethylacetate=20:1˜5:1) to afford compound C85-12 (1.2 g, light yellow solid,yield 53.6%). MS m/z (ESI): 373.8 [M+H]⁺.

Step 12:

Guanidine hydrochloride (920 mg, 0.0096 mmol) was dissolved in anhydrousethanol (20 mL). Under the protection of nitrogen, sodium methoxide(0.864 g, 0.016 mol) was added, the reaction was stirred at roomtemperature for half an hour, and then compound C85-12 (1.2 g, 0.0032mol) was added. The reaction solution was heated to reflux, and stirredfor 18 hours. LC-MS indicated the reaction of the starting materials wascomplete. The reaction solution was cooled to room temperature,filtered, the filtrate was concentrated under reduced pressure, and thecrude product was purified by column chromatography on silica gel (DCM:MeOH=50:1˜20:1) to afford compound C85 (500 mg, light yellow solid,yield 46.7%).

¹H NMR (400 MHz, DMSO) δ 8.19 (s, 1H), 7.91 (s, 1H), 6.71 (s, 2H), 6.58(s, 1H), 6.54 (s, 2H), 3.15˜3.19 (m, 1H), 1.31 (d, J=6.8 Hz, 6H). MS m/z(ESI): 339.7 [M+H]⁺.

The compound in the following table was prepared according to a methodsimilar to that described in Example 24.

Starting material or regent different from Compound that in Example No.Compound Structure Name 24 Characterization Data C84 (compound 84)

5-((2-chloro-5- isopropylpyridin- 4- yl)thio)pyrimidine- 2,4-diamineHydrobromic acid and liquid bromine in Step 3 was replaced withconcentrated hydrochloric acid; aniline hydrobromide in Step 11 wasreplaced with ¹H NMR (400 MHz, CD₃OD) δ 8.12 (s, 1H), 7.94 (s, 1H), 6.12(s, 1H), 3.35- 3.33 (m, 1H), 1.37 (d, J = 6.8 Hz, 6H). MS m/z (ESI):295.8 [M + H]⁺. aniline hydrochloride.

Example 25: preparation of5-((5-isopropyl-2-vinylpyridine-4-yl)thio)pyrimidine-2,4-diamine (C95,compound 95) and5-((2-ethyl-5-isopropylpyridin-4-yl)thio)pyrimidine-2,4-diamine (C127,compound 127)

Step 1:

Compound C85 (50 mg, 0.15 mol) was dissolved in 1,4-dioxane (10 mL) andwater (1 mL), vinylboronic acid (47 mg, 0.3 mmol), K₂CO₃ (42 mg, 0.3mol) and Pd(PPh₃)₄ (12 mg) were sequentially added. Under the protectionof nitrogen, the reaction was performed at 100° C. for 18 hours. LC-MSindicated the reaction of the starting materials was substantiallycomplete, and the target product was obtained. The reaction solution wascooled to room temperature, and filtered. The filter cake was washedwith 1,4-dioxane (10 mL), and concentrated under reduced pressure todryness. The crude product was purified by column chromatography onsilica gel (dichloromethane:methanol=50:1-10:1) to afford compound C95(15 mg, white solid, yield 34.8%).

¹H NMR (400 MHz, CD₃OD) δ 8.27 (s, 1H), 7.97 (s, 1H), 6.79 (s, 1H), 6.67(dd, J=17.6, 11.2 Hz, 1H), 5.93 (d, J=17.6 Hz, 1H), 5.42 (d, J=11.2 Hz,1H), 3.42-3.3 (n, 1H), 1.38 (d, J=6.8 Hz, 6H). MS m/z (ESI): 287.9[M+H]S.

Step 2:

Compound C95 (10 mg, 0.03 mol) was dissolved in anhydrous methanol (5mL), 10% palladium/carbon (5 mg) was added, and the reaction wasperformed under hydrogen (0.4 MPa) at room temperature for 18 hours.LC-MS indicated the reaction of the starting materials was complete. Thereaction solution was filtered, the filter cake was washed with 10 mlmethanol, concentrated under reduced pressure, and purified by columnchromatography on silica gel (dichloromethane:methanol=10:1) to affordcompound C127 (3 mg, white solid, yield 29.8%).

¹H NMR (400 MHz, CD₃OD) δ 8.20 (s, 1H), 7.94 (s, 1H), 6.60 (s, 1H),3.39-3.30 (m, 1H), 2.64 (d, J=7.6 Hz, 2H), 1.39 (d, J=6.8 Hz, 6H), 1.17(t, J=7.6 Hz, 3H). MS m/z (ESI): 289.9 [M+H]7.

The compounds in the following table were prepared according to methodssimilar to that described in Example 25.

Starting material or regent different Compound Compound from that No.Structure Name in Example 25 Characterization Data C92 (compound 92)

5-((5- isopropyl-2- methylpyridin- 4-yl)thio) pyrimidine-2,4- diamineVinylboronic acid in Step 1 was replaced with methylboronic acid, andStep 2 was omitted. ¹H NMR (400 MHz, CD₃OD) δ 8.18 (s, 1H), 7.95 (s,1H), 6.60 (s, 1H), 3.39-3.29 (m, 1H), 2.37 (s, 3H), 1.38 (d, J = 6.8 Hz,6H). MS m/z (ESI): 275.8 [M + H]⁺. C109 (compound 109)

5-((2-(1- ethoxyethyl)-5- isopropyl- pyridin-4- yl)oxy) pyrimidine-2,4-diamine The preparation started from Step 2, and C95 in Step 2 wasreplaced with C45. ¹H NMR (400 MHz, CD₃OD) δ 8.51 (s, 1H), 8.02 (s, 1H),7.51 (s, 1H), 4.81-4.76 (m, 1H), 3.62-3.52 (m, 3H), 1.48 (d, J = 5.2 Hz,3H), 1.41 (d, J = 6.0 Hz, 6H), 1.23 (t, J = 6.8 Hz, 3H). MS m/z (ESI):317.9 [M + H]⁺. C91 (compound 91)

5-((5- isopropyl2- (methyltluo) pyridin-4- yl)thio) pyrimidine-2,4-diamine Vinylboronic acid in Step 1 was replaced with sodiumthiomethoxide, Pd(PPh₃)₄ was replaced with Pd₂(dba)₃ and Xantphos, K₂CO₃was replaced with DIEA, and Step 2 was omitted. ¹H NMR (400 MHz, CD₃OD)δ 8.25 (s, 1H), 8.15 (s, 1H), 6.67 (s, 1H), 3.36-3.27 (m, 1H), 2.50 (s,3H), 1.38 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 307.8 [M + H]⁺. C125(compound 125)

5-((2- (ethylthio)-5- isopropyl- pyridin-4- yl)thio) pyrimidine-2,4-diamine Vinylboronic acid in Step 1 was replaced with sodiumethanethiolate, Pd(PPh₃)₄ was replaced with Pd₂(dba)₃ and Xantphos,K₂CO₃ was replaced with DIEA. ¹H NMR (400 MHz, CD₃OD) δ 8.16 (s, 1H),7.94 (s, 1H), 6.51 (s, 1H), 3.34-3.28 (m, 1H), 3.01-2.88 (m, 2H),1.38-1.31 (m, 3H). 1.27 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 321.9 [M +H]⁺. C165 (compound 165)

5-((5-ethyl-2- methoxy- pyridin-4- yl)oxy) pyrimidine- 2,4-diamine Thepreparation started from Step 2, and C95 in Step 2 was replaced withC162. ¹H NMR (400 MHz, CD₃OD) δ 7.88 (s, 1H), 5.97 (s, 1H), 5.51 (s,1H), 3.84 (s, 3 H). 2.75- 2.73 (m, 2H), 1.32-1.22 (m, 3H). MS m/z (ESI):261.9 [M + H]⁺. C194 (compound 194)

5-((2- cyclopropyl-5- isopropyl- pyridin-4- yl)oxy)-N⁴- isobutyl-pyrimidine- 2,4-diamine C85 in Step 1 was replaced with C181;vinylboronic acid was replaced with cyclopropyl- boronic acid, and Step2 was omitted. ¹H NMR (400 MHz, CD₃OD) δ 8.39 (s, 1H), 7.86 (s, 1H),7.09 (s, 1H), 3.50-3.46 (m, 1H), 3.37-3.33 (m, 2H), 2.27 (m, 1H), 2.03(m, 1H), 1.41-1.33 (m, 8H), 1.14 (s, 2H), 0.94 (m, 6H). MS m/z (ESI):341.9 [M + H]⁺.

Example 26: preparation of5-((2-bromo-5-isopropylpyridin-4-yl)oxy)-N²-methylpyrimidine-2,4-diamine(C237, compound 237)

Step 1:

Compound C2-5 (5 g, 0.023 mol) was dissolved in acetone (200 mL), ethylbromoacetate (5.8 g, 0.035 mol), and potassium carbonate (6.4 g, 0.046mol) were sequentially added, and the reaction solution was stirred at50° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was quenched by adding water (100mL), and extracted with ethyl acetate (100 mL×3). The organic phaseswere combined, washed once with saturated brine (100 mL), then driedover anhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated to afford a crude product, compound C237-1 (5 g crudeproduct, brown oily liquid, yield 74%), which was directly used in thenext step. MS m/z (ESI): 288.0 [M+H]⁺.

Step 2:

C237-1 (30 g, 0.091 mol) was dissolved in 1,2-dimethoxyethane (100 mL),and ethyl formate (10 g, 0.14 mol) and potassium tert-butoxide (20 g,0.18 mol) were added to the solution. The reaction was performed at 25°C. for 1 hour. LC-MS indicated the reaction was complete. The reactionsolution was poured into petroleum ether (100 mL), a large amount ofwhite solid precipitated, which was filtered and dried to afford C237-2(35 g, white solid, yield 95.6%). MS m/z (ESI): 329.9 [M+H]⁺.

Step 3:

C237-2 (35 g, 0.11 mol) was dissolved in ethanol (100 mL), thiourea(24.3 g, 0.32 mol) and sodium methoxide (17.3 g, 0.32 mol) were added tothe solution. The reaction was performed at 100° C. overnight. LC-MSindicated the reaction was complete. The reaction solution was rotaryevaporated to dryness to afford a yellow residue, which was purified bycolumn chromatography on silica gel (petroleum ether:ethyl acetate=1:1)to afford compound C237-3 (5 g, yellow solid, yield 13.8%). MS m/z(ESI): 341.7 [M+H]⁺.

Step 4:

C237-3 (5 g, 0.015 mol) was added to water (20 mL), sodium hypochlorite(10 mL) and sodium hydroxide (1.2 g) were added to the solution. Thereaction was performed at 90° C. for 20 hours. LC-MS indicated thereaction was complete. The reaction was adjusted to pH 7 with 1M HCl,and extracted with ethyl acetate (100 mL×3). The organic phases werecombined, washed once with saturated brine (100 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated. The crude product was isolated by column chromatography onsilica gel (petroleum ether:ethyl acetate=1:1) to afford compound C237-4(3 g, light yellow solid, yield 63%). MS m/z (ESI): 325.7 [M+H]⁺.

Step 5:

Phosphorus oxybromide (10 g) was added to C237-4 (3 g, 9.23 mmol), andthe reaction was performed at 70° C. for 2 hours. LC-MS indicated thereaction was complete. The reaction solution was slowly added to icewater, adjusted to pH 7 with an aqueous solution of sodium bicarbonate,then extracted with ethyl acetate (50 mL×3). The organic phases werecombined, washed once with saturated brine (50 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated. The crude product was isolated by column chromatography onsilica gel (petroleum ether:ethyl acetate=10:1) to afford compoundC237-5 (2 g, light yellow solid, yield 48.3%). MS m/z (ESI): 449.4[M+H]⁺.

Step 6:

C237-5 (1.8 g, 0.004 mol) was added to DCM (10 mL), and aqueous ammonia(20 ml) was added to the solution. The reaction was performed at 40° C.for 4 hours. LC-MS indicated the reaction was complete. The reaction wasextracted with ethyl acetate (50 mL×3), the organic phases werecombined, washed once with saturated brine (40 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated. The crude product was isolated by column chromatography onsilica gel (petroleum ether:ethyl acetate=1:1) to afford compound C237-6(1.2 g, white solid, yield 77.4%). MS m/z (ESI): 386.5 [M+H]⁺.

Step 7:

Compound C237-6 (100 mg, 0.26 mmol) was dissolved in NMP (3 mL),methylamine hydrochloride (20 mg) was added to the solution, and thereaction was performed under microwave radiation at 150° C. for 1 hour.The reaction solution was directly purified by preparative liquidchromatography to afford C237 (30 mg, white solid, yield 34.4%).

¹H NMR (400 MHz, CD₃OD) δ 8.24 (s, 1H), 7.82 (s, 1H), 7.12 (s, 1H),3.34-3.32 (m, 1H), 3.02 (s, 3H), 1.37 (d, J=7.2 Hz, 6H). MS m/z (ESI):337.9 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 26.

Starting material or regent different Compound Compound from that in No.Structure Name Example 26 Characterization Data C111 (compound 111)

2-((4-amino-5- ((2-bromo-5- isopropyl- pyridin-4- yl)oxy) pyrimidin-2-yl)amino) propane-1,3-diol Phosphorus oxybromide in Step 5 was replacedwith phosphorus oxychloride; methylamine hydrochloride in Step 7 wasreplaced with serinol. ¹H NMR (400 MHz, CD₃OD) δ 8.16 (s, 1H), 7.65 (s,1H), 6.76 (s, 1H), 4.05 (m, 1H), 3.74 (d, J = 5.6 Hz, 4H), 3.36- 3.33(m, 1H), 1.38 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 397.8 [M + H]⁺. C218(compound 218)

2-((4-amino-5- ((2-chloro-5- isopropyl- pyridin-4- yl)oxy) pyrimidin-2-yl)amino) propane-1,3-diol C2-5 in Step 1 was replaced with 2- chloro-5-isopropylpyridin- 4-ol; phosphorus oxybromide in Step 5 was replacedwith phosphorus oxychloride; methylamine hydrochloride in ¹H NMR (400MHz, CD₃OD) δ 8.18 (s, 1H), 7.65 (s, 1H), 6.64 (s, 1H), 4.04 (m, 1H),3.73 (d, J = 5.2 Hz, 4H), 3.43- 3.39 (m, 1H), 1.38 (d, J = 6.8 Hz, 6H).MS m/z (ESI): 353.8 [M + H]⁺. Step 7 was replaced with serinol. C235(compound 235)

3-((4-amino-5- ((2-bromo-5- isopropyl- pyridin-4- yl)oxy) pyrimidin-2-yl)amino) propane- 1,2-diol Methylamine hydrochloride in Step 7 wasreplaced with 3- aminopropane-1,2- diol. ¹H NMR (400 MHz, DMSO- d₆) δ8.65 (s, 1H), 8.24 (m, 2H), 8.03 (s, 1H), 7.72 (s, 1H), 7.25 (s, 1H),3.67 (m, 1H), 3.45-3.22 (m, 7H), 1.27 (d, J = 6.8, 6H). MS m/z (ESI):397.7 [M + H]⁺. C243 (compound 243)

3-((4-((4-amino- 2-((2,3- dihydroxypropyl) amino) pyrimidin-5-yl)oxy)-5- isopropylpyridin- 2-yl)amino) propane-1,2-diol Methylaminehydrochloride in Step 7 was replaced with 3- aminopropane-1,2- diol. ¹HNMR (400 MHz, CD₃OD) δ 7.98 (s, 1H), 7.68 (s, 1H), 6.48 (s, 1H),3.88-3.83 (m, 2H), 3.61-3.28 (m, 9H), 1.33 (d, J = 6.8 Hz, 6H). MS m/z(ESI): 408.6 [M + H]⁺. C236 (compound 236)

2-((4-amino-5- ((2-bromo-5- isopropyl- pyridin-4- yl)oxy) pyrimidin-2-yl)amino)ethanol Methylamine hydrochloride in Step 7 was replaced withethanolamine. ¹H NMR (400 MHz, DMSO- d₆) δ 8.63 (s, 1H), 8.24-8.18 (m,2H), 8.03-7.94 (m, 2H), 7.25 (s, 1H), 3.60-3.57 (m, 2H), 3.55-3.34 (m,2H), 3.32- 3.26 (m, 1H), 1.27 (d, J = 7.2 Hz, 6H). MS m/z (ESI): 367.9[M + H]⁺. C242 (compound 242)

2-((4-((4-amino- 2-((2- hydroxyethyl) amino) pyrimidin-5- yl)oxy)-5-isopropyl- pyridin-2- yl)amino)ethanol Methylamine hydrochloride in Step7 was replaced with ethanolamine. ¹H NMR (400 MHz, CD₃OD) δ 7.98 (s,1H), 7.67 (s, 1H), 6.43 (s, 1H), 3.79-3.75 (m, 4H), 3.47-3.45 (m, 2H),3.34- 3.33 (m, 2H), 3.31-3.28 (m, 1H), 1.33 (d, J= 7.2 Hz, 6H). MS m/z(ESI): 348.8 [M + H]⁺. C238 (compound 238)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N²- cyclopropyl-pyrimidine-2,4- diamine Methylamine hydrochloride in Step 7 was replacedwith cyclopropylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.25 (s, 1H), 7.81 (s,1H), 7.16 (s, 1H), 3.42-3.38 (m, 1H), 2.74 (m, 1H), 1.38 (d, J = 6.8 Hz,6H), 0.95 (m, 2H), 0.74 (m, 2H). MS m/z (ESI): 363.9 [M + H]⁺. C239(compound 239)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)-N²- isopropyl- pyrimidine-2,4-diamine Methylamine hydrochloride in Step 7 was replaced withisopropylamine. ¹H NMR (400 MHz, CD₃OD) δ 8.24 (s, 1H), 7.78 (s, 1H),7.14 (s, 1H), 4.24-4.22 (m, 1H), 3.41-3.38 (m, 1H), 1.37 (d, J = 6.8 Hz,6H), 1.31 (d, J = 6.4 Hz, 6H). MS m/z (ESI): 367.9 [M + H]⁺.

Example 27: preparation of5-((5-isopropyl-2-(methylselanyl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C121, compound 121)

Step 1:

Compound C2 (50 mg, 0.15 mol) was dissolved in anhydrous DMF (5 mL),sodium hydride (18 mg, 0.75 mmol) was added under ice bath cooling, thereaction was stirred for 30 minutes, followed by addition ofp-methoxybenzyl chloride (118 mg, 0.75 mmol). Under the protection ofnitrogen, the reaction was performed at room temperature for 2 hours.LC-MS indicated the reaction of the starting materials was substantiallycomplete, and the target product was obtained. The reaction solution wasslowly dropwise added with water (5 mL), and extracted with ethylacetate (10 ml×3). The organic phases were combined, added with 10 ganhydrous sodium sulfate, dried for 30 min, and filtered. The filtercake was washed with ethyl acetate (10 mL), and concentrated underreduced pressure to dryness. The crude product was purified by columnchromatography on silica gel (mobile phase: petroleum ether:ethylacetate=20:1˜5:1), to afford compound C121-1 (20 mg, oil, yield 16.5%).MS m/z (ESI): 339.8 [M+H]⁺.

Step 2:

Compound C121-1 (20 mg, 0.025 mmol) was dissolved in anhydroustetrahydrofuran (5 mL), the reaction was cooled to −78° C., a solutionof n-butyl lithium (2.4 mol/L, 0.04 ml) was slowly dropwise added underthe protection of nitrogen, and the reaction was stirred at thistemperature for 15 minutes. Dimethyldiselenide was added, the reactionwas stirred at this temperature for 15 minutes, then allowed to slowlywarm to room temperature, and the reaction was performed at roomtemperature for 2 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete, and the target product wasobtained. The reaction solution was slowly dropwise added with 5 mlwater, and extracted with ethyl acetate (12 ml×3). The organic phaseswere combined, added with 10 g anhydrous sodium sulfate, dried for 30min, and filtered. The filter cake was washed with ethyl acetate (10mL), the filtrate was concentrated, and then purified by columnchromatography on silica gel (mobile phase: petroleum ether:ethylacetate=20:1˜3:1), to afford compound C121-2 (5 mg, oil, yield 24.4%).MS m/z (ESI): 819.6 [M+H]⁺.

Step 3:

Compound C121-2 (4 mg, 0.005 mmol) was dissolved in trifluoroacetic acid(1 mL), the reaction was stirred at room temperature for 1 hour, andthen warmed to 45° C. and stirred for 2 hours. LC-MS indicated thereaction of the starting materials was substantially complete, and thetarget product was obtained. The solvent was concentrated under reducedpressure to dryness, and the crude product was purified by thin layerchromatography on silica gel (DCM:MeOH=10:1) to afford compound C121(1.12 mg, white solid, yield 66.1%).

¹H NMR (400 MHz, CD₃OD) δ 8.61 (s, 1H), 8.21 (s, 1H), 7.42 (s, 1H),3.08-3.12 (m, 1H), 2.48 (s, 3H), 1.38 (d, J=6.8 Hz, 6H). MS m/z (ESI):339.8 [M+H]⁺.

Example 28: preparation ofN⁴-(2-(dimethylamino)ethyl)-5-((5-isopropyl-2-(methylthio)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C72, compound 72)

Compound C71 (12 mg, 0.031 mmol) was dissolved in 1,4-dioxane (5 mL),sodium thiomethoxide (13 mg, 0.19 mmol), DIEA (40 mg, 0.31 mmol), X-phos(5 mg) and Pd₂(dba)₃ (5 mg) were sequentially added, and the reactionwas stirred at 90° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was substantially complete. The reaction solution wasconcentrated under reduced pressure to afford a crude product, which waspurified by thin layer chromatography on silica gel(dichloromethane:methanol=10:1), to afford compound C72 (2 mg, whitesolid, yield 18%).

¹H NMR (400 MHz, CD₃OD) δ 8.26 (s, 1H), 7.60 (s, 1H), 6.54 (s, 1H),3.81-3.78 (m, 2H), 3.36-3.35 (m, 1H), 2.95 (s, 6H), 2.48 (s, 3H), 1.37(d, J=7.2 Hz, 6H), 0.92-0.90 (m, 2H). MS m/z (ESI): 362.9 [M+H]⁺.

Example 29: preparation ofN-(4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpyridin-2-yl)acetamide(C122, compound 122)

Compound C21 (25 mg, 0.096 mmol) was dissolved in dichloromethane (5mL), acetic anhydride (29 mg, 0.29 mmol) and triethylamine (29 mg, 0.29mmol) were sequentially added, and the reaction solution was stirred at25° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete. The reaction solution wasconcentrated under reduced pressure to afford a crude product, which waspurified by thin layer chromatography on silica gel(dichloromethane:methanol=10:1), to afford compound C122 (10 mg, whitesolid, yield 34%).

¹H NMR (400 MHz, DMSO-d₆) δ 10.34 (s, 1H), 8.06 (s, 1H), 7.52 (s, 1H),7.41 (s, 1H), 6.31 (s, 2H), 5.95 (s, 2H), 2.60 (s, 1H), 1.91 (s, 3H),1.27 (d, J=6.8 Hz, 6H). MS m/z (ESI): 302.9 [M+H]⁺.

Example 30: preparation of4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpicolinic acid (C128,compound 128)

C54 (10 mg, 0.037 mmol) was dissolved in ethanol (5 mL) and water (1mL), and sodium hydroxide (3 mg, 0.074 mmol) was added. The reaction wasstirred at 80° C. for 3 hours. LC-MS indicated the reaction wascomplete. Then, the reaction was adjusted to pH 7 with 0.5N hydrochloricacid, concentrated to dryness, and the residue was purified by thinlayer chromatography on silica gel (dichloromethane:methanol=10:1), toafford C128 (3 mg, white solid, yield 28.0%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.59 (s, 1H), 8.19 (s, 2H), 7.98 (s, 1H),7.68 (s, 2H), 7.34 (s, 1H), 3.60-3.56 (m, 1H), 1.33 (d, J=6.8 Hz, 6H).MS m/z (ESI): 289.9 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 30.

Starting material or regent different Compound Compound from that in No.Structure Name Example 30 Characterization Data C197 (compound 197)

4-((2,4- diaminopyrimidin- 5-yl)oxy)-6- methoxynicotinamide C54 wasreplaced with C177. ¹H NMR (400 MHz, CD₃OD) δ 8.53 (s, 1H), 8.51 (s,1H), 7.54 (s, 1H), 3.99 (s, 3H). MS m/z (ESI): 277.0 [M + H]⁺. C198(compound 198)

4-((2,4- diaminopyrimidin-5- yl)oxy)-6- methoxynicotinic acid C54 wasreplaced with C177; and the duration of stirring was extended to 15hours. ¹H NMR (400 MHz, CD₃OD) δ 8.46 (s, 1H), 7.73 (s, 1H), 6.23 (s,1H), 3.92 (s, 3H). MS m/z (ESI): 278.1 [M + H]⁺.

Example 31: preparation of4-((2,4-diaminopyrimidin-5-yl)oxy)-5-isopropylpicolinimidamide (C131,compound 131)

Step 1:

C54 (20 mg, 0.074 mmol) was dissolved in methanol (5 mL), and sodiummethoxide (8 mg, 0.15 mmol) was added. The reaction was stirred at roomtemperature overnight. LC-MS indicated the reaction was complete. Then,the reaction was concentrated to dryness to give C131-1 (20 mg, oil,yield 89.3%). MS m/z (ESI): 302.9 [M+H]⁺.

Step 2

C131-1 (10 mg, 0.033 mmol) was dissolved in ethanol (5 mL) and water (2mL), and ammonium chloride (2 mg, 0.039 mmol) was added. The reactionwas stirred at 80° C. for 2 hours. LC-MS indicated the reaction wascomplete. Then, the reaction was concentrated to dryness, and theresidue was purified by preparative thin layer chromatography on silicagel (dichloromethane:methanol=10:1) to afford C131 (6 mg, white solid,yield 63.1%).

¹H NMR (400 MHz, DMSO-d₆) δ 9.47 (m, 1H), 8.66 (s, 1H), 8.25 (s, 1H),8.01 (s, 1H), 7.84 (s, 2H), 7.70 (s, 1H), 3.43-3.40 (m, 1H), 1.33 (d,J=6.8, 6H). MS m/z (ESI): 287.9 [M+H]⁺.

Example 32: preparation ofN⁵-(2-ethyl-5-isopropylpyridin-4-yl)pyrimidin-2,4,5-triamine (C132,compound 132)

Step 1:

C85-7 (10 g, 0.0467 mol) was dissolved in 1,4-dioxane (100 mL),vinylpinacol borate (14.4 g, 0.093 mmol), potassium carbonate (19 g,0.14 mol) and Pd(PPh₃)₄ (1 g, 0.865 mmol) were sequentially added. Purgewith nitrogen was performed for 3 times, and the reaction was performedat 90° C. overnight. LC-MS indicated the reaction was complete. Then,the reaction solution was poured into water (200 mL), and extracted withethyl acetate (200 mL×3). The organic phases were combined, washed oncewith saturated brine (200 mL), then dried over anhydrous sodium sulfate(50 g) for half an hour, filtered, and concentrated.

The crude product was purified by column chromatography on silica gel(petroleum ether:ethyl acetate=1:1) to afford compound C132-2 (6 g,yellow oil, yield 79.2%). MS m/z (ESI): 163.0 [M+H]⁺.

Step 2:

C132-2 (6 g, 0.037 mol) was dissolved in ethanol (100 mL), 10% wetpalladium/carbon (2 g) was added, and the reaction was performed underhydrogen at room temperature for 2 hours. LC-MS indicated the reactionwas complete. Then, the reaction was filtered, and concentrated to giveC132-3 (4.5 g, yellow oil, yield 74.1%). MS m/z (ESI): 165.1 [M+H]⁺.

Step 3:

C132-3 (4.5 g, 0.03 mol) was dissolved in acetonitrile (50 mL), Boc₂O(7.3 g, 0.033 mol), DMAP (4 g, 0.033 mol) and triethylamine (6 g, 0.06mol) were sequentially added. The reaction was stirred at roomtemperature for 6 hours. LC-MS indicated the reaction was complete.Then, the reaction was quenched by adding water, and extracted withethyl acetate (50 mL×3). The organic phases were combined, washed oncewith saturated brine (50 mL), then dried over anhydrous sodium sulfate(20 g) for half an hour, filtered, and concentrated. The crude productwas purified by column chromatography on silica gel (petroleumether:ethyl acetate=1:1) to afford compound C132-4 (1.4 g, white solid,yield 19.4%). MS m/z (ESI): 265.0 [M+H]⁺.

Step 4:

C132-4 (1.4 g, 5.28 mmol) was dissolved in DMF (10 mL), sodium hydride(0.38 g, 15.8 mmol) and bromoacetonitrile (1.26 g, 10.6 mmol) weresequentially added, and the reaction was stirred at room temperatureovernight. LC-MS indicated the reaction was complete. Then, the reactionwas quenched by adding water, and extracted with ethyl acetate (100mL×3). The organic phases were combined, washed with saturated brine (40mL×3), then dried over anhydrous sodium sulfate (50 g) for half an hour,filtered, and concentrated. The crude product was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=10:1) toafford compound C132-5 (1 g, oil, yield 62.5%). MS m/z (ESI): 304.0[M+H]⁺.

Step 5:

C132-5 (1 g, 3.3 mmol) was dissolved in DMF (10 mL), and tert-butoxybis(dimethylamino)methane (1.7 g, 9.9 mmol) was added to the reactionsolution. The reaction was stirred at 100° C. for 1 hour. LC-MSindicated the reaction was complete. Then, the reaction was quenched byadding water, and extracted with ethyl acetate (30 mL×3). The organicphase was concentrated, and the residue was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=5:1) toafford C132-6 (0.5 g, yield 37.6%). MS m/z (ESI): 359.0 [M−45+H]*.

Step 6:

C132-6 (200 mg, 0.56 mmol) was dissolved in DMF (1 mL), and anilinehydrobromide (110 mg, 0.62 mmol) was added to the reaction solution. Thereaction was stirred at 100° C. for 6 hours. LC-MS indicated thereaction was complete. Then, the reaction was quenched by adding water,extracted with ethyl acetate, concentrated, and the residue was purifiedby column chromatography on silica gel (petroleum ether:ethylacetate=5:1) to afford compound C132-7 (80 mg, yield 39.8%). MS m/z(ESI): 407.0 [M+H]⁺.

Step 7:

C132-7 (20 mg, 0.049 mmol) was dissolved in ethanol (2 mL), andguanidine hydrochloride (100 mg, 1.05 mmol) and sodium methoxide (30 mg,0.55 mmol) were added to the reaction solution. The reaction was stirredat 90° C. for 6 hours. LC-MS indicated the reaction was complete. Then,the reaction was concentrated to dryness, and the residue was purifiedby Prep-HPLC to afford C132 (5 mg, yield 37.3%).

¹H NMR (400 MHz, CD₃OD) δ 8.06 (s, 1H), 7.92 (s, 1H), 6.73 (s, 1H),3.21-3.18 (m, 1H), 2.83-2.79 (m, 2H), 1.39 (d, J=6.8 Hz, 6H), 1.32-1.28(m, 3H). MS m/z (ESI): 273.0 [M+H]⁺.

Example 33: preparation of5-((5-isopropyl-2-(trimethylsilyl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C133, compound 133)

Compound C2 (200 mg, 0.62 mmol) was dissolved in tetrahydrofuran (10mL), and n-butyl lithium (1 mL, 2.4 mmol) was dropwise added under theprotection of nitrogen at −78° C. The reaction solution was stirred at−78° C. for half an hour, and then bromotrimethylsilane (1 mL) wasdropwise added. The reaction solution was stirred at −78° C. for 2hours. LC-MS indicated the reaction of the starting materials wassubstantially complete. The reaction solution was quenched by addingwater, adjusted to pH 7 with sodium carbonate, extracted with ethylacetate (25 mL×3), dried over sodium sulfate, and concentrated underreduced pressure to afford a crude product. The crude product was rinsedwith ethyl acetate (2 mL), filtered and dried to afford compound C133 (5mg, yellow solid, yield 2.5%).

¹H NMR (400 MHz, CDCl₃) δ 8.69 (s, 1H), 7.76 (s, 1H), 6.88 (s, 1H),3.44-3.40 (m, 1H), 1.39 (d, J=7.2 Hz, 6H), 0.35 (s, 9H). MS m/z (ESI):317.9 [M+H]⁺.

Example 34: preparation of5-((2-(sec-butyl)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C141, compound 141)

Step 1:

C2-4 (5.0 g, 22 mmol), tributyl(1-ethoxyvinyl)tin (8.7 g, 24 mmol) anddioxane (40 mL) were added to a 500 mL single-neck flask, Pd(PPh₃)₂Cl₂(1.5 g, 2.2 mmol) and cuprous iodide (420 mg, 2.2 mmol) were added,purge with argon was performed for 3 times, and the reaction was placedin an oil bath at 100° C. and allowed to proceed for 1.5 h. After thereaction was complete, the reaction was cooled to room temperature,filtered with suction, and the filtrate was concentrated under reducedpressure. The residue was purified by column chromatography on silicagel (petroleum ether:ethyl acetate=10:1), to afford C141-2 (4.64 g,light yellow oil, yield 96.6%/o).

¹H NMR (400 MHz, CDCl₃) δ 8.28 (s, 1H), 7.17 (s, 1H), 5.35 (d, J=2.0 Hz,1H), 4.33 (d, J=2.0 Hz, 1H), 3.98 (q, J=7.0 Hz, 2H), 3.90 (s, 3H),3.25-3.18 (m, 1H), 1.44 (t, J=7.0 Hz, 3H), 1.24 (d, J=7.0 Hz, 6H). MSm/z (ESI): 222.0 [M+H]⁺.

Step 2:

C141-2 (4.64 g, 21 mmol) was dissolved in dichloromethane (40 mL), a 4mol/L solution of hydrochloric acid in dioxane (15.6 mL, 63 mmol) wasadded at room temperature, and the reaction was performed at roomtemperature for 20 h. After the reaction was complete, the reaction wasconcentrated under reduced pressure, adjusted to pH 6˜7 with a 2 mol/Laqueous solution of sodium hydroxide, and extracted with ethyl acetate(100 mL×3). The organic phases were combined, then washed with asaturated aqueous solution of sodium chloride (100 mL×2), dried overanhydrous sodium sulfate, filtered, and the filtrate was concentrated toafford C141-3 (4.0 g, yellow oil, yield 99.6%).

¹H NMR (400 MHz, CDCl₃) δ 8.39 (s, 1H), 7.54 (s, 1H), 3.93 (s, 3H),3.33-3.23 (m, 1H), 2.70 (s, 3H), 1.27 (d, J=6.8 Hz, 6H). MS m/z (ESI):194.0 [M+H]⁺.

Step 3:

At 0° C., to a suspension of ethyltriphenylphosphonium bromide (10.7 g,29 mmol) in anhydrous tetrahydrofuran (80 mL), a 2.4 mol/L solution ofbutyllithium in n-hexane (12.1 mL, 29 mmol) was dropwise added, and thereaction was performed at 0° C. for 1 h. A solution of C141-3 (3.7 g, 19mmol) in tetrahydrofuran (20 mL) was dropwise added at 0° C. to theabove suspension, and after the dropwise addition, the reaction waswarmed to room temperature and performed for 18 hours. After thereaction was complete, the reaction system was quenched by dropwiseaddition of a saturated aqueous solution of ammonium chloride (100 mL),and extracted with ethyl acetate (100 mL×2). The organic phases werecombined, then washed with a saturated aqueous solution of sodiumchloride (100 mL×2), dried over anhydrous sodium sulfate, and filtered.The filtrate was concentrated, and the residue was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=6:1) toafford C141-4 (2.02 g, colorless oil, yield 51.5%).

¹H NMR (400 MHz, CDCl₃) δ 8.26 (s, 1H), 6.82 (s, 1H), 6.38-6.33 (m, 1H),3.88 (s, 3H), 3.24-3.14 (m, 1H), 2.09 (s, 3H), 1.84 (dd, J=6.8, 0.8 Hz,3H), 1.24 (d, J=6.8 Hz, 6H). MS m/z (ESI): 206.0 [M+H]⁺.

Step 4:

C141-4 (1.9 g, 9.25 mmol) was dissolved in ethanol (100 mL), 10%palladium/carbon (1.9 g, 100% wt) was added, purge with hydrogen wasperformed for 3 times with a hydrogen balloon, and the reaction wasperformed at room temperature for 18 h. After the reaction was complete,the reaction was filtered with suction, and the filtrate wasconcentrated to afford C141-5 (1.3 g, colorless oil, crude product,yield 67.8%). MS m/z (ESI): 208.1 [M+H]⁺.

Step 5:

C141-5 (1.3 g, 6.3 mmol) was dissolved in dichloromethane (39 mL), andboron tribromide (5 mL) was dropwise added at room temperature. Afterthe dropwise addition, the reaction was placed in an oil bath at 45° C.and allowed to proceed for 18 h. After the reaction was complete, thereaction solution was poured into ice water, adjusted to pH 7 with a 2mol/L aqueous solution of sodium hydroxide, and extracted with ethylacetate (50 mL×4). The organic phases were combined, then washed with asaturated aqueous solution of sodium chloride (100 mL×2), dried overanhydrous sodium sulfate, filtered, and the filtrate was concentratedunder reduced pressure to afford C141-6 (1.3 g, off-white solid, crudeproduct, yield 100%). MS m/z (ESI): 194.0 [M+H]⁺.

Step 6:

C141-6 (500 mg, 2.59 mmol) was dissolved in anhydrous DMF (5 mL),anhydrous potassium carbonate (716 mg, 5.18 mmol) and2-bromoacetonitrile (372 mg, 3.1 mmol) were sequentially added to thesolution, and the reaction was performed at room temperature for 8 h.The reaction solution was filtered, the filtrate was diluted with ethylacetate (100 mL), then washed with a saturated aqueous solution ofsodium chloride (100 mL×2), dried over anhydrous sodium sulfate, andfiltered. The filtrate was concentrated under reduced pressure, and theresidue was purified by column chromatography on silica gel (petroleumether:ethyl acetate=3:1) to afford C141-7 (380 mg, light yellow oil,yield 63.2%).

¹H NMR (400 MHz, CDCl₃) δ 8.35 (s, 1H), 6.61 (s, 1H), 4.85 (s, 2H),3.24-3.14 (m, 1H), 2.82-2.74 (m, 1H), 1.81-1.70 (m, 1H), 1.68-1.57 (m,1H), 1.28 (d, J=7.6 Hz, 3H), 1.27 (d, J=6.8 Hz, 6H), 0.86 (t, J=7.4 Hz,3H). MS m/z (ESI): 233.1 [M+H]⁺.

Step 7:

C141-7 (600 mg, 2.6 mmol) and tert-butoxy bis(dimethylamino)methane(1.36 g, 7.8 mmol) was dissolved in anhydrous DMF (10 mL), and thereaction was placed in an oil bath at 100° C. and allowed to proceed for2 h. After the reaction was complete, the reaction was cooled to roomtemperature, diluted with ethyl acetate (40 mL), then washed with asaturated aqueous solution of sodium chloride (50 mL×2), dried overanhydrous sodium sulfate, and filtered. The filtrate was concentrated,and then the residue was purified by column chromatography on silica gel(petroleum ether:ethyl acetate=2:1) to afford C141-8 (320 mg, yellowoil, crude product, yield 37.2%). MS m/z (ESI): 287.9 [M−45+H]*.

Step 8:

C141-6 (21 mg, 0.073 mmol) and aniline hydrobromide (38 mg, 0.22 mmol)were dissolved in anhydrous ethanol (5 mL), and the reaction was placedin an oil bath at 80° C. and allowed to proceed for 2 h. After thereaction was complete, the reaction was cooled to room temperature, thereaction system was added with sodium methoxide (60 mg, 1.11 mmol) andguanidine hydrochloride (60 mg, 0.62 mmol), and the reaction was againplaced in an oil bath at 80° C. and allowed to proceed for 16 h. Afterthe reaction was complete, the reaction was cooled to room temperature,filtered with suction, the filtrate was concentrated, and the residuewas purified by preparative high-performance liquid chromatography toafford compound C141 (3.25 mg, off-white solid, yield 17.0%).

¹H NMR (300 MHz, DMSO-d₆) δ 8.29 (s, 1H), 7.52 (s, 1H), 6.35 (s, 2H),6.29 (s, 1H), 5.97 (s, 2H), 3.38 (m, 1H), 2.61 (m, 1H), 1.63-1.53 (m,1H), 1.49-1.40 (m, 1H), 1.29 (d, J=6.9 Hz, 6H), 1.10 (d, J=6.9 Hz, 3H),0.73 (t, J=7.2 Hz, 3H). MS m/z (ESI): 302.1 [M+H]⁺.

Example 35: preparation of2,2′-((5-((2-bromo-5-isopropylpyridin-4-yl)oxy)pyrimidin-2,4-diyl)bis(azanediyl))diethanol(C226, compound 226)

Compound C237-5 (50 mg, 0.11 mmol) was dissolved in N-methylpyrrolidone(5 mL), ethanolamine (0.1 g) was added, the reaction solution wasstirred under microwave radiation at 150° C. for 1 hour. LC-MS indicatedthe reaction of the starting materials was substantially complete. Thereaction solution was concentrated under reduced pressure to afford acrude product, which was purified by preparative liquid chromatographyto afford compound C226 (3 mg, white solid, yield 7%).

¹H NMR (400 MHz, CD₃OD) δ 8.22 (s, 1H), 7.68 (s, 1H), 7.01 (s, 1H),3.77-3.72 (m, 4H), 3.66-3.65 (m, 2H), 3.64-3.58 (m, 2H), 3.43-3.40 (m,1H), 1.37 (d, J=6.8 Hz, 6H). MS m/z (ESI): 411.7 [M+H]⁺.

The compound in the following table was prepared according to a methodsimilar to that described in Example 35.

Starting material or regent different Compound Compound from that in No.Structure Name Example 35 Characterization Data C227 (compound 227)

5-((2-bromo-5- isopropylpyridin- 4-yl)oxy)- N²,N⁴- dimethylpyrimidine-2,4-diamine Ethanolamine in Step 1 was replaced with methylamine. ¹H NMR(400 MHz, CD₃OD) δ 8.25 (s, 1H), 7.72 (s, 1H), 7.13 (s, 1H), 3.41-3.38(m, 1H), 3.08-3.06 (m, 6H), 1.37 (d, J = 7.2 Hz, 6H). MS m/z (ESI):351.7 [M + H]⁺.

Example 36: preparation of5-((2-((2,4-dimethoxyphenyl)ethynyl)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C144, compound 144)

Step 1:

Compound C66 (20 mg, 0.074 mmol) was dissolved in DMF (5 mL),2,4-dimethoxybromobenzene (48 mg, 0.22 mmol), TEA (38 mg, 0.37 mmol),CuI (10 mg) and Pd(PPh₃)₂Cl₂ (10 mg) were sequentially added, purge withnitrogen was performed for 3 times, and the reaction was performed at50° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete. The reaction solution was cooledto room temperature, filtered, the filtrate was added with purifiedwater (50 mL), and extracted with ethyl acetate (50 mL×3). The organicphases were combined, added with anhydrous sodium sulfate (10 g), driedfor 30 min, filtered, and concentrated under reduced pressure to afforda crude product, which was purified by Prep-HPLC to afford compound C144(0.9 mg, yield 3.0%).

¹H NMR (400 MHz, CD₃OD) δ 8.45 (s, 1H), 7.84 (s, 1H), 7.47 (d, J=8.4 Hz,1H), 7.25 (s, 1H), 6.62-6.58 (m, 2H), 3.90 (s, 3H), 3.86 (s, 3H),3.47-3.33 (m, 1H), 1.41 (d, J=6.8 Hz, 6H). MS m/z (ESI): 405.9 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 36.

Starting material or regent different from that in No. CompoundStructure Compound Name Example 36 Characterization Data C145 (compound145)

5-((5-isopropyl-2- ((2,4,6- trimethoxyphenyl) ethynyl)pyridin-4-yl)oxy)pyrimidine- 2,4-diamine 2,4- dimethoxybromo- benzene was replacedwith 2,4,6- trimethoxybromo- benzene. ¹H NMR (400 MHz, CD₃OD) δ 8.36 (s,1H), 7.72 (s, 1H), 6.93 (s, 1H), 6.25 (s, 2H), 3.86 (s, 9H), 3.50-3.33(m, 1H), 1.40 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 435.8 [M + H]⁺. C149(compound 149)

5-((5-isopropyl-2- (quinolin-8- ylethynyl)pyridin- 4- yl)oxy)pyrimidine-2,4-diamine 2,4- dimethoxybromo- benzene was replaced with 8-bromoquinoline. ¹H NMR (400 MHz, CD₃OD) δ 8.95 (d, J = 4.4 Hz, 1H),8.50- 8.40 (m, 2H), 8.12 (d, J = 7.2 Hz, 1H), 8.04 (d, J = 8.4 Hz, 1H),7.70-7.60 (m, 3H), 7.33 (s, 1H), 3.54-3.50 (m, 1H), 1.44 (d, J = 6.8 Hz,6H). MS m/z (ESI): 396.8 [M + H]⁺. C150 (compound 150)

5-((5-isopropyl-2- (pyridin-2- ylethynyl)pyridin- 4- yl)oxy)pyrimidine-2,4-diamine 2,4- dimethoxybromo- benzene was replaced with 2-bromopyridine. ¹H NMR (400 MHz, CD₃OD) δ 8.73-8.70 (m, 2H), 8.10-8.03(m, 2H), 7.91-7.89 (m, 1H), 7.82 (s, 1H), 7.68-7.65 (m, 1H), 3.59-3.55(m, 1H), 1.45 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 346.9 [M + H]⁺. C151(compound 151)

5-((5-isopropyl-2- (pyridin-4- ylethynyl)pyridin- 4- yl)oxy)pyrimidine-2,4-diamine 2,4- dimethoxybromo- benzene was replaced with 4-bromopyridine. ¹H NMR (400 MHz, CDCl₃) δ 8.65 (d, J = 5.2 Hz, 2H), 8.52(s, 1H), 7.71 (s, 1H), 7.45 (d, J = 5.2 Hz, 2H), 6.91 (s, IH), 5.60 (s,2H), 5.10 (s, 2H), 3.43- 3.39 (m, 1H), 1.41 (d, J = 6.8 Hz, 6H). MS m/z(ESI): 346.9 [M + H]⁺. C195 (compound 195)

5-((2- (imidazo[1,2- b]pyridazin-3- ylethynyl)-5- isopropylpyridin- 4-yl)oxy)pyrimidine- 2,4-diamine 2,4- dimethoxybromo- benzene was replacedwith 3- bromoimidazo [1,2- b]pyridazine. ¹H NMR (400 MHz, CD₃OD) δ 8.63(s, 1H), 8.42 (s, 1H), 8.13- 8.10 (m, 2H), 7.66 (s, 1H), 7.39- 7.37 (m,1H), 6.95 (s, 1H), 3.53-3.48 (m, 1H), 1.43 (d, J = 6.8 Hz, 6H). MS m/z(ESI): 386.9 [M + H]⁺.

Example 37: preparation of5-((2-(aminomethyl)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C157, compound 157) and5-((2-((dimethylamino)methyl)-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C219, compound 219)

Step 1:

C54 (100 mg, 0.37 mmol) was dissolved in tetrahydrofuran (10 mL), andlithium aluminum hydride (84 mg, 2.22 mmol) was added at 0° C. Thereaction was stirred at 0° C. for 30 minutes. LC-MS indicated thereaction was complete. Then, the reaction was quenched by adding water(1 mL), and filtered. The filtrate was concentrated to dryness, and theresidue was purified by the Prep-HPLC method to afford compound C157(100 mg, yellow solid, yield 98.5%).

¹H NMR (400 MHz, CD₃OD) δ 8.55 (s, 1H), 7.73 (s, 1H), 7.03 (s, 1H), 4.20(s, 2H), 3.48-3.45 (m, 1H), 1.39 (d, J=7.2 Hz, 6H). MS m/z (ESI): 274.9[M+H]⁺.

Step 2:

C157 (60 mg, 0.22 mmol) was dissolved in ethanol (10 mL), and an aqueoussolution of formaldehyde (17.7 mg, 0.22 mmol) and benzotriazole (26 mg,0.22 mmol) were sequentially added. The reaction was stirred at roomtemperature for 6 hours, followed by addition of sodium borohydride(41.5 mg, 1.10 mmol), and then the reaction was stirred at roomtemperature for 1 hour. LC-MS detected the product was formed. Thereaction was filtered, the filtrate was concentrated to dryness, and theresidue was purified by the Prep-HPLC method to afford C219 (1 mg, whitesolid, yield 1.5%).

¹H NMR (400 MHz, CD₃OD) δ 8.59 (s, 1H), 7.77 (s, 1H), 7.05 (s, 1H), 4.37(s, 2H), 3.49-3.46 (m, 1H), 2.92 (s, 6H), 1.40 (d, J=6.8 Hz, 6H). MS m/z(ESI): 302.8 [M+H]⁺.

Example 38: preparation of5-((2-(ethylamino)-5-(prop-1-en-2-yl)pyridin-4-yl)oxy)pyrimidine-2,4-diamine(C214, compound 214)

Step 1:

Compound C21-2 (10 g, 0.061 mol) was dissolved in tert-butanol (30 mL),di-tert-butyl dicarbonate (13 g, 0.073 mol) was added, and the reactionsolution was stirred at 30° C. for 16 hours. LC-MS indicated thereaction of the starting materials was substantially complete. Then, thereaction was added with purified water (150 mL), and extracted withethyl acetate (200 mL×3). The organic phases were combined, washed withan aqueous solution of NaCl (100 mL), added with anhydrous sodiumsulfate (20 g), dried for 30 min, filtered, and concentrated underreduced pressure to afford a crude product, which was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=10:1˜5:1) toafford compound C214-2 (10 g, yellow oily liquid, yield 62%). MS m/z(ESI): 265.1 [M+H]⁺.

Step 2:

Compound C214-2 (10 g, 0.038 mol) was dissolved in DMF (30 mL), sodiumhydride (1.8 g, 0.076 mol) was added, and the reaction solution wasstirred at 0° C. for 15 minutes. Then, iodoethane (8.9 g, 0.057 mmol)was added, and the reaction solution was stirred at 0° C. for 1 hour.LC-MS indicated the reaction of the starting materials was substantiallycomplete. Then, then reaction was added with purified water (150 mL),and extracted with ethyl acetate (200 mL×3). The organic phases werecombined, washed with an aqueous solution of NaCl (100 mL), added withanhydrous sodium sulfate (20 g), dried for 30 min, filtered, andconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=10:1˜5:1) to afford compound C214-3 (5.5 g, light yellow oilyliquid, yield 55%). MS m/z (ESI): 292.9 [M+H]⁺.

Step 3:

Compound C214-3 (5 g, 0.026 mol) was dissolved in N-methylpyrrolidone(20 mL), p-toluenesulfonic acid (22.5 g, 0.13 mol) and lithium chloride(5.5 g, 0.13 mol) were sequentially added, and the reaction solution wasstirred at 180° C. for 1 hour. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was directly usedin the next step. MS m/z (ESI): 179.0 [M+H]⁺.

Step 4:

DMF (10 mL) was added to the reaction solution of compound C214-4, thenbromoacetonitrile (6.9 g, 0.056 mol) and potassium carbonate (39 g, 0.28mol) were sequentially added, and the reaction solution was stirred atroom temperature for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction was quenched by addingwater (100 mL), and extracted with ethyl acetate (100 mL×3). The organicphases were combined, washed once with saturated brine (100 mL), thendried over anhydrous sodium sulfate (30 g) for half an hour, filtered,and concentrated under reduced pressure to afford a crude product, whichwas purified by column chromatography on silica gel (petroleumether:ethyl acetate=5:1˜1:1) to afford compound C214-5 (4 g, brown oilyliquid, yield 67%). MS m/z (ESI): 218.0 [M+H]⁺.

Step 5:

Compound C214-5 (3.5 g, 0.016 mol) was dissolved in DMF (20 mL),tert-butoxy bis(dimethylamino)methane (8.4 g, 0.048 mol) was added, andthe reaction solution was stirred at 100° C. for 1 hour. LC-MS indicatedthe reaction of the starting materials was complete. The reactionsolution was quenched by adding water (100 mL), and extracted with ethylacetate (100 mL×3). The organic phases were combined, washed withsaturated brine (50 mL×3), then dried over anhydrous sodium sulfate (20g) for half an hour, filtered, and concentrated under reduced pressureto afford a crude product, which was purified by column chromatographyon silica gel (petroleum ether:ethyl acetate=5:1˜1:1) to afford compoundC214-6 (2.3 g, brown oily liquid, yield 45%). MS m/z (ESI): 273.1[M+H]⁺.

Step 6:

Compound C214-6 (2.3 g, 7.32 mmol) was dissolved in ethanol (10 mL),aniline hydrobromide (1.9 g, 0.011 mol) was added, and the reactionsolution was stirred at 100° C. for 16 hours. LC-MS indicated thereaction of the starting materials was complete. The reaction solutionwas cooled to room temperature, quenched by adding water (50 mL), andextracted with ethyl acetate (50 mL×3). The organic phases werecombined, then washed with saturated brine (50 mL), added with anhydroussodium sulfate (10 g), dried for half an hour, and filtered. Thefiltrate was concentrated under reduced pressure to afford a crudeproduct, which was purified by column chromatography on silica gel(petroleum ether:ethyl acetate=1:1), to afford compound C214-7 (800 mg,red solid, yield 35%). MS m/z (ESI): 321.1 [M+H]⁺.

Step 7:

Compound C214-7 (100 mg, 0.31 mmol) was dissolved in ethanol (5 mL),guanidine hydrochloride (89 mg, 0.93 mmol) and sodium methoxide (50 mg,0.93 mmol) were sequentially added, and the reaction was stirred at 90°C. for 16 hours. LC-MS indicated the reaction of the starting materialswas complete. The reaction solution was cooled to room temperature, andconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (ethyl acetate) toafford compound C214 (24 mg, light yellow solid, yield 24%).

¹H NMR (400 MHz, CD₃OD) δ 7.80 (s, 1H), 7.57 (s, 1H), 5.83 (s, 1H), 5.17(s, 1H), 5.13 (s, 1H), 3.22 (q, J=6.8 Hz, 2H), 2.17 (s, 3H), 1.18 (t,J=7.2 Hz, 3H). MS m/z (ESI): 286.8 [M+H]⁺.

The compound in the following table was prepared according to methodssimilar to that described in Example 38.

Starting material or regent different from that in Characterization No.Compound Structure Compound Name Example 38 Data C192 (compound 192)

5-((2- (methylamino)-5- (prop-1-en-2- yl)pyridin-4- yl)oxy)pyrimidine-2,4-diamine Iodoethane in Step 2 was replaced with iodomethane. ¹H NMR(400 MHz, CD₃OD) δ 7.82 (s, 1H), 7.58 (s, 1H), 5.86 (s, 1H), 5.18 (s,1H), 5.15 (s, 1H), 2.82 (s, 3H), 2.17 (s, 3H). MS m/z (ESI): 272.8 [M +H]⁺.

Example 39: preparation of5-((2-methoxy-5-(prop-1-en-2-yl)pyridin-4-yl)methyl)pyrimidine-2,4-diamine(C196, compound 196)

Step 1:

Compound C196-1 (10 g, 0.053 mol) was dissolved in anhydroustetrahydrofuran (50 mL), lithium diisopropylamide (32 mL, 0.064 mol) wasadded at −65° C., and the reaction solution was stirred at −65° C. for15 minutes. Then, DMF (5.8 g, 0.080 mol) was dropwise added, and thereaction solution was stirred at −65° C. for 30 minutes. LC-MS indicatedthe reaction of the starting materials was substantially complete.

The reaction solution was poured into water (100 mL), and extracted withethyl acetate (200 mL×3). The organic phases were combined, washed withsaturated brine (100 mL), dried over anhydrous sodium sulfate (20 g) forhalf an hour, filtered, and concentrated under reduced pressure toafford a crude product, which was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=100:1), to afford compoundC196-2 (4 g, yellow oily liquid, yield 36%). MS m/z (ESI): 215.8 [M+H]⁺.

Step 2:

Compound C196-2 (4 g, 0.019 mol) was dissolved in 1,4-dioxane (50 mL),and isopropenylpinacol borate (6.25 g, 0.072 mol), potassium carbonate(7.7 g, 0.056 mol), Pd(PPh₃)₄ (0.1 g) and water (5 mL) were sequentiallyadded. Purge with nitrogen was performed for 3 times, and the reactionsolution was stirred at 100° C. for 18 hours. LC-MS indicated thereaction of the starting materials was complete. The reaction solutionwas cooled to room temperature, and filtered. The filtrate wasconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=10:1), to afford compound C196-3 (2.4 g, yellow oily liquid,yield 73%). MS m/z (ESI): 178.0 [M+H]⁺.

Step 3

Compound C196-3 (354 mg, 0.0020 mol) was dissolved in methanol (10 mL),acrylonitrile (212 mg, 0.0040 mol), sodium methoxide (212 mg, 0.0030mol) were sequentially added, and the reaction solution was stirred atroom temperature for 16 hours. LC-MS indicated the reaction of thestarting materials was substantially complete. The reaction wasfiltered, and concentrated under reduced pressure to afford a crudeproduct C196-4, and the crude product was directly used in the next step(350 mg, light yellow oily liquid, yield 64%). MS m/z (ESI): 205.9[M+H]⁺.

Step 4:

Compound C196-4 (300 mg, 1.09 mmol) was dissolved in methanol (20 mL),guanidine carbonate (1.3 g, 10.9 mmol) was added, and the reactionsolution was stirred at 80° C. for 16 hours. LC-MS indicated thereaction of the starting materials was complete. The reaction solutionwas filtered, and the filtrate was concentrated under reduced pressureto afford a crude product, which was purified by column chromatographyon silica gel (petroleum ether:ethyl acetate=5:1) to afford compoundC196 (150 mg, white solid, yield 51%).

¹H NMR (400 MHz, DMSO-d) δ 7.96 (s, 1H), 7.23 (s, 1H), 6.50 (s, 1H),5.26 (s, 1H), 4.91 (s, 1H), 3.82 (s, 3H), 3.61 (s, 2H), 1.99 (s, 3H). MSm/z (ESI): 272.0 [M+H]⁺.

Example 40: preparation ofN⁴-(2-aminoethyl)-5-((2-bromo-5-isopropylpyridin-4-yl)oxy)pyrimidine-2,4-diamine(C199, compound 199)

Step 1:

Compound C237-1 (4.5 g, 0.015 mol) was dissolved in DMF (30 mL), DMF-DMA(5.6 g, 0.047 mol) was added, and the reaction solution was stirred at130° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was complete. The reaction was quenched by adding water (100mL), and extracted with ethyl acetate (100 mL×3). The organic phaseswere combined, washed once with saturated brine (100 mL), then driedover anhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=10:1˜5:1) to afford compound C199-1 (2 g, yellow oily liquid,yield 40%). MS m/z (ESI): 342.8 [M+H]⁺.

Step 2:

Compound C199-1 (2 g, 0.0056 mol) was dissolved in ethanol (20 mL),guanidine hydrochloride (3.2 g, 0.034 mol) and sodium methoxide (1.82 g,0.034 mol) were sequentially added, and the reaction solution wasstirred at 90° C. for 16 hours. LC-MS indicated the reaction of thestarting materials was complete. The reaction solution was filtered, thefiltrate was concentrated under reduced pressure, added with purifiedwater (50 mL), then adjusted to a pH value of 7 with 1M HCl, andextracted with ethyl acetate (100 mL×3). The organic phases werecombined, washed once with saturated brine (100 mL), then dried overanhydrous sodium sulfate (10 g) for half an hour, filtered, andconcentrated to afford a crude product, which was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=20:1˜5:1) toafford compound C199-2 (0.7 g, light yellow solid, yield 38%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.19 (s, 1H), 7.88 (s, 1H), 7.67 (s, 2H),7.16 (s, 1H), 3.25-3.18 (m, 1H), 1.27 (d, J=6.8 Hz, 6H). MS m/z (ESI):324.8 [M+H]⁺.

Step 3:

Compound C199-2 (200 mg, 0.617 mmol) was dissolved in phosphorusoxychloride (1 mL), the reaction was heated to 80° C., and stirred for16 hours. LC-MS indicated the reaction of the starting materials wascomplete. The reaction solution was cooled to room temperature, quenchedby adding water (30 mL), then adjusted to a pH value of 7-8 with sodiumbicarbonate, and extracted with dichloromethane (50 mL×3). The organicphases were combined, then washed with saturated brine (50 mL), addedwith anhydrous sodium sulfate, dried for half an hour, and filtered. Thefiltrate was concentrated under reduced pressure, and the crude productwas purified by column chromatography on silica gel (petroleumether:ethyl acetate=10:1˜1:1), to afford compound C199-3 (90 mg, lightyellow solid, yield 42%).

¹H NMR (400 MHz, CDCl₃) δ 8.24 (s, 1H), 8.16 (s, 1H), 6.58 (s, 1H), 5.35(s, 2H), 3.38-3.35 (m, 1H), 1.38 (d, J=6.8 Hz, 6H). MS m/z (ESI): 342.7[M+H]⁺.

Step 4:

Compound C199-3 (15 mg, 0.044 mmol) was dissolved in acetonitrile (5mL), tert-butyl (2-aminoethyl)carbamate (0.5 mL) was added, and thereaction solution was stirred at 90° C. for 16 hours. LC-MS indicatedthe reaction of the starting materials was substantially complete. Thereaction solution was concentrated under reduced pressure to afford acrude product C199-4, and this crude product was directly used in thenext step. (10 mg, light yellow oily liquid, yield 47%). MS m/z (ESI):467.0 [M+H]⁺.

Step 5:

Compound C199-4 (10 mg, 0.021 mmol) was dissolved in dichloromethane (5mL), trifluoroacetic acid (1 mL) was added, and the reaction solutionwas stirred at room temperature for 30 minutes. LC-MS indicated thereaction of the starting materials was substantially complete. Thereaction solution was concentrated under reduced pressure to afford acrude product, which was purified by preparative high-performance liquidchromatography to afford compound C199 (4 mg, white solid, yield 50%).

¹H NMR (400 MHz, CD₃OD) δ 8.26 (s, 1H), 7.80 (s, 1H), 7.21 (s, 1H), 3.84(t, J=5.6 Hz, 6H), 3.42-3.37 (m, 1H), 3.25 (t, J=5.6 Hz, 2H), 1.37 (d,J=7.2 Hz, 6H). MS m/z (ESI): 366.7 [M+H]⁺.

The compound in the following table was prepared according to a methodsimilar to that described in Example 40.

Starting material or regent different Compound from that in No. CompoundStructure Name Example 40 Characterization Data C217 (compound 217)

5-((2-bromo- 5- isopropylpyridin- 4-yl)oxy)-4- hydrazineylpyrimidin-2-amine Tert-butyl (2- aminoethyl)carbamate in Step 4 was replaced withtert- butyl hydrazinecarboxylate. ¹H NMR (400 MHz, CD₃OD) δ 8.50 (s,1H), 7.72 (s, 1H), 7.38 (s, 1H), 2.98-2.92 (m, 1H), 1.32 (d, J = 6.8 Hz,6H). MS m/z (ESI): 338.8 [M + H]⁺.

Example 41: preparation of5-((2-ethynyl-5-isopropylpyridin-4-yl)oxy)-N⁴-isopropylpyrimidine-2,4-diamine(C212, compound 212)

Step 1:

Compound C199-3 (700 mg, 0.020 mol) was dissolved in anhydroustetrahydrofuran (20 mL), trimethylsilylacetylene (200 mg, 0.020 mol),N,N-diisopropylethylamine (1.2 g, 0.0095 mol), cuprous iodide (40 mg,0.21 mmol) and Pd(PPh₃)₂Cl₂ (143 mg, 0.21 mmol) were sequentially added,and the reaction solution was stirred under the protection of nitrogenat 50° C. for 2 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete. The reaction solution wasconcentrated under reduced pressure to afford a crude product, which waspurified by column chromatography on silica gel (petroleum ether:ethylacetate=10:1˜5:1), to afford compound C212-1 (400 mg, yellow solid,yield 55%). MS m/z (ESI): 361.1 [M+H]⁺.

Step 2:

Compound C212-1 (20 mg, 0.056 mmol) was dissolved in acetonitrile (5mL), isopropylamine was added, and the reaction solution was stirred at90° C. for 16 hours. LC-MS indicated the reaction of the startingmaterials was substantially complete. The reaction solution wasconcentrated under reduced pressure to afford a crude product, which waspurified by preparative thin layer chromatography on silica gel(dichloromethane:methanol=10:1), to afford compound C212-2 (15 mg,yellow oily liquid, yield 71%). MS m/z (ESI): 383.8 [M+H]⁺.

Step 3:

Compound C212-2 (15 mg, 0.039 mmol) was dissolved in methanol (5 mL),potassium carbonate (5 mg) was added, and the reaction solution wasstirred at room temperature for 20 minutes. LC-MS indicated the reactionof the starting materials was complete. The reaction solution wasfiltered, and the filtrate was concentrated under reduced pressure togive a crude product, which was purified by preparative thin layerchromatography on silica gel (dichloromethane:methanol=10:1), to affordcompound C212 (3 mg, white solid, yield 25%).

¹H NMR (400 MHz, CD₃OD) δ 8.36 (s, 1H), 7.52 (s, 1H), 6.75 (s, 1H),4.42-4.39 (m, 1H), 3.70 (s, 1H), 3.50-3.45 (m, 1H), 1.38 (d, J=7.2 Hz,6H), 1.21 (d, J=6.4 Hz, 6H). MS m/z (ESI): 311.8 [M+H]⁺.

The compounds in the following table were prepared according to methodssimilar to that described in Example 41.

Starting material or regent different Compound from that in No.Structure Compound Name Example 41 Characterization Data C213 (compound213)

5-((2-ethynyl-5- isopropylpyridin- 4-yl)oxy)-N⁴- methylpyrimidine-2,4-diamine Isopropylamine in Step 2 was replaced with methylamine. ¹HNMR (400 MHz, CD₃OD) δ 8.36 (s, 1H), 7.54 (s, 1H), 6.76 (s, 1H), 3.70(s, 1H), 3.45 (m, 1H), 2.94 (s, 3H), 1.39 (d, J = 6.8 Hz, 6H). MS m/z(ESI): 283.8 [M + H]⁺. C211 (compound 211)

2-((2-amino-5-((2- ethynyl-5- isopropylpyridin- 4- yl)oxy)pyrimidin-4-yl)amino)ethanol Isopropylamine in Step 2 was replaced withethanolamine. ¹H NMR (400 MHz, CD₃OD) δ 8.34 (s, 1H), 7.54 (s, 1H), 6.78(s, 1H), 3.69-3.67 (m, 3H), 3.57-3.54 (m, 2H), 3.49- 3.46 (m, 1H), 1.39(d, J = 6.8 Hz, 6H). MS m/z (ESI): 313.9 [M + H]⁺. C216 (compound 216)

3-((2-amino-5-((2- ethynyl-5- isopropylpyridin- 4- yl)oxy)pyrimidin- 4-yl)amino)propane- 1,2-diol Isopropylamine in Step 2 was replaced with 3-aminopropane- 1,2-diol. ¹H NMR (400 MHz, CD₃OD) δ 8.35 (s, 1H), 7.57 (s,1H), 6.80 (s, 1H), 3.81-3.79 (m, 1H), 3.70 (s, 1H), 3.53-3.44 (m, 5H),1.39 (d, J = 6.8 Hz, 6H). MS m/z (ESI): 343.8 [M + H]⁺.

Reference Example: preparation of5-((5-chloro-2-isopropylpyridin-3-yl)oxy)pyrimidine-2,4-diamine (00,compound 10)

Step 1:

At 20° C., to compound C10-1 (20 g, 0.155 mol) in a 10% aqueous solutionof NaOH (140 mL), Br₂ (24.5 g, 0.153 mol) in a 10% aqueous solution ofNaOH (144 mL) was slowly dropwise added. The reaction was stirred atthis temperature for 30 min, and then the reaction solution was slowlypoured into water (300 mL). The large amount of white solid thusobtained was filtered to afford compound C10-2 (15 g, crude product),and the crude product was directly used in the next step. MS m/z (ESI):207.8 [M+H]⁺.

Step 2:

At 25° C., compound C10-2 (17 g, 0.07 mol), bromoacetonitrile (17.5 g,0.1 mol) were dissolved in DMF (100 mL), and then K₂CO₃ (19.3 g, 0.14mol) was slowly added. The reaction mixture was stirred at thistemperature overnight. The reaction solution was slowly poured intowater (500 mL). The aqueous phase was extracted with ethyl acetate, theorganic phase was washed with saturated brine, dried over anhydroussodium sulfate, filtered, and concentrated under reduced pressure togive a crude product. The crude product was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=10:1,100˜200 mesh silica gel) to afford compound C10-3 (8.5 g, light yellowsolid, yield: 49.36%).

¹H NMR (400 MHz, DMSO-d₆) δ 8.23 (s, 1H), 7.94 (s, 1H), 5.38 (s, 2H); MSm/z (ESI): 246.7 [M+H]⁺.

Step 3:

At 25° C. and under the protection of nitrogen,tetrakis(triphenylphosphine)palladium(0.8 g) was added to a mixed systemof compound C10-3 (8.5 g, 0.034 mol), isopropenylpinacol borate (11.6 g,0.069 mol), water (10 mL), potassium carbonate (9.6 g, 0.069 mol) and1,4-dioxane (200 mL). The reaction was stirred at 90° C. overnight. Thereaction solution was poured into water (300 mL). The aqueous phase wasextracted with ethyl acetate, the organic phase was washed withsaturated brine, dried over anhydrous sodium sulfate, filtered,concentrated under reduced pressure to give a crude product. The crudeproduct was purified by column chromatography on silica gel (petroleumether:ethyl acetate=10:1, 100˜200 mesh silica gel) to afford compoundC10-4 (5.5 g, crude product, light yellow oil). MS m/z (ESI): 208.9[M+H]⁺.

Step 4:

At 20° C., wet Pd/C (0.5 g) was added to a solution of compound C10-4(5.5 g, 0.0259 mol) in ethanol (50 mL). Purge with hydrogen wasperformed 3 times, the reaction was performed under hydrogen (0.4 Mpa),and then the reaction mixture was filtered. The filtrate wasconcentrated to dryness to afford compound C10-5 (5.2 g, light brownoil, crude product). The crude product was directly used in the nextstep. MS m/z (ESI): 171.9 [M+H]⁺.

Step 5:

At 25° C., K₂CO₃ (8.4 g, 0.06 mol) was slowly added to a solution ofcompound C10-5 (5.2 g, 0.03 mol) and bromoacetonitrile (5.5 g, 0.046mol) in DMF (50 mL). The reaction was stirred at this temperature for 16hours. The reaction solution was poured into water (300 mL). The aqueousphase was extracted with ethyl acetate, separated, the organic phase waswashed with saturated brine, dried over anhydrous sodium sulfate,filtered, and concentrated under reduced pressure to give a crudeproduct. The crude product was purified by column chromatography onsilica gel (petroleum ether:ethyl acetate=30:1, 100˜200 mesh silica gel)to afford compound C10-6 (2.7 g, oil product, crude product). MS m/z(ESI): 210.9 [M+H]⁺.

Step 6:

At 25° C., tert-butoxy bis(dimethylamino)methane (1 mL) was added to asolution of compound C10-6 (1 g, 4.76 mmol) in DMF (5 mL). The reactionwas stirred at 100° C. for 1 h. The reaction solution was poured intowater (50 mL). The aqueous phase was extracted with ethyl acetate, theorganic phase was washed with saturated brine, dried over anhydroussodium sulfate, filtered, and concentrated under reduced pressure togive a crude product. The crude product was purified by columnchromatography on silica gel (petroleum ether:ethyl acetate=5:1, 100˜200mesh silica gel) to afford compound C10-7 (0.84 g, oil, crude product).MS m/z (ESI): 265.9 [M+H]⁺.

Step 7:

At 25° C., aniline hydrochloride (0.2 g, 1.5 mmol) was added to asolution of compound C10-7 (0.4 g, 1.3 mmol) in DMF (2 mL). The reactionwas stirred at 100° C. for 5 hours. TLC (petroleum ether:ethylacetate=5:1, UV) indicated the reaction was complete. The reactionsolution was poured into water (20 mL). The aqueous phase was extractedwith ethyl acetate, the organic phase was washed with saturated brine,dried over anhydrous sodium sulfate, filtered, and concentrated underreduced pressure to give a crude product. The crude product was purifiedby column chromatography on silica gel (petroleum ether:ethylacetate=20:1, 100˜200 mesh silica gel) to afford C10-8 (0.26 g, lightyellow solid, yield: 64.00%).

Step 8:

At 25° C., guanidine carbonate (155 mg, 1.27 mmol) was added to asolution of compound C10-8 (200 mg, 0.64 mmol) in ethanol (10 mL). Thereaction was stirred under microwave radiation at 100° C. for 1 h. Thereaction solution was concentrated under reduced pressure to give acrude product. The crude product was added to a solution of petroleumether:ethyl acetate=1:30, stirred for 1 h, and filtered to affordcompound C10 (20 mg, light yellow solid, yield: 11.2%, compound 10).

¹H NMR (400 MHz, DMSO-d₆) δ 8.23 (s, 1H), 7.58 (s, 1H), 6.90 (s, 1H),6.48 (s, 2H), 6.00 (s, 2H), 3.53-3.49 (m, 1H), 1.26 (d, J=6.8 Hz, 6H);MS m/z (ESI): 279.9 [M+H]⁺.

The following compounds were prepared according to methods similar tothose described in the above Examples.

No. Structure MS Data C24

303.9 C25

318.9 C28

353.8 C29

267.8 C32

276.9 C36

353.9 C37

328.9 C50

301.9 C51

301.9 C53

313.9 C55

303.9 C56

289.9 C57

289.9 C58

315.9 C59

315.9 C60

301.9 C62

316.9 C64

316.9 C68

313.9 C69

394.0 C70

350.0 C73

278.9 C74

322.8 C75

370.8 C76

274.9 C77

268.9 C78

323.9 C79

322.9 C80

290.9 C81

258.9 C82

312.9 C83

312.9 C86

387.8 C87

291.9 C89

340.8 C90

339.9 C93

329.9 C94

329.9 C96

301.9 C97

270.9 C98

284.9 C99

269.9 C100

283.9 C101

277.9 C102

321.8 C103

369.8 C104

289.9 C105

321.9 C106

322.9 C107

273.9 C108

267.9 C110

349.9 C112

266.9 C113

283.8 C114

265.9 C123

314.9 C130

320.8 C139

377.9 C140

372.9 C146

377.9 C147

393.9 C156

288.9 C158

303.9 C159

304.9 C160

292.9 C161

305.9 C166

333.9 C167

405.8 C168

413.8 C169

337.8 C178

275.9 C184

359.9 C185

349.9 C186

343.0 C187

399.0 C188

383.1 C189

329.9 C190

313.9 C191

283.9 C193

325.9 C200

299.9 C206

301.9 C220

318.9 C221

287.9 C223

298.9 C224

360.9 C225

326.9 C232

309.9 C234

384.9 C240

270.9 C244

330.9 C245

322.9 C247

341.9 C248

338.9

Biological Assay: Determination of the Inhibitory Activity of theCompounds on Human P2X3 and P2X2/3 Receptors

Cells were seeded into a poly-D-lysine-coated 384-well cell cultureplate (Corning) at a density of 11,000 cells/well/25 μL of cellinoculation medium, and were cultured in a cell incubator overnight. Onthe day of the test, the calcium 6 dye was diluted to a 2× concentrationwith a assay buffer, 25 μL of the 2× calcium 6 dye was added to the384-well cell culture plate, which was incubated at 37° C. for 2 hours,and then placed at room temperature for further use. The test compoundand the agonist, α,β-MeATP were diluted to a 7× concentration with theassay buffer, 10 μL of the 7× test compound was added to the 384-wellcell culture plate, which was incubated at room temperature for 15minutes, and 10 μL of the 7× α,β-MeATP was transferred into the 384-wellcell culture plate. The data were measured and analyzed using FLIPRTetra, and the half inhibitory concentration (IC₅₀) of the test compoundon P2X3 and P2X2/3 receptors was calculated with the GraphPad Prismfour-parameter equation.

Cell lines: human embryonic kidney cells HEK293-P2X3 and HEK293-P2X2/3stably expressing cell lines;

Complete cell culture medium: DMEM High Glucose (Life Technology), whichcontained 10% fetal bovine serum, 4 mM GlutaMAX, 1%penicillin-streptomycin, and 350 μg/mL G418;

Cell inoculation medium: DMEM High Glucose (Life Technology), whichcontained 2% fetal bovine serum, and 4 mM GlutaMAX;

Cell culture conditions: 37° C., 5% CO₂;

Assay buffer: HBSS (containing calcium and magnesium ions), whichcontained 20 mM HEPES;

Detection equipment: FLIPR Tetra (Molecular Devices);

Detection parameters: excitation wavelength 470-495 nm, emissionwavelength 515-575 nm; fluorescence signal was measured once everysecond for 260 seconds in total.

The experimental data obtained from the above biological assay are shownin the table below.

P2X3 Compound No. IC₅₀ (μM) C1 0.7460 C2 0.0881 C3 0.2047 C4 0.1022 C60.348 C9 0.2543 Reference Compound CIO 2.5070 C12 0.0736 C13 0.3552 C150.3070 C16 0.8015 C20 0.0444 C21 0.131 C22 0.037 C23 0.3650 C33 0.238C35 1.1670 C38 0.787 C39 0.166 C41 1.8540 C43 0.0550 C44 0.1156 C450.4350 C52 0.147 C61 0.274 C65 0.334 C66 0.085 C67 0.490 C71 0.072 C720.127 C84 0.536 C85 0.078 C88 0.133 C91 0.227 C92 0.109 C95 0.138 C1110.104 C117 0.151 C118 0.142 C119 0.540 C120 0.492 C124 1.243 C125 0.731C127 0.147 C129 0.377 C133 1.441 C134 0.391 C135 0.303 C136 0.270 C1370.503 C138 0.635 C142 0.302 C145 0.809 C148 0.215 C149 0.130 C150 0.211C151 0.59 C152 0.211 C153 0.083 C154 0.072 C155 0.926 C157 0.736 C1620.296 C164 0.046 C165 0.163 C172 0.116 C173 0.188 C174 0.100 C175 0.108C176 0.611 C179 0.051 C180 0.207 C181 0.084 C182 0.67 C192 0.053 C1940.322 C195 0.316 C196 0.208 C199 0.186 C201 0.316 C202 0.103 C203 0.157C204 0.044 C207 0.113 C208 0.07528 C209 0.144 C210 0.269 C211 0.064 C2120.081 C213 0.112 C214 0.044 C215 0.08818 C216 0.062 C218 0.186 C2220.048 C226 0.1146 C227 0.948 C228 0.119 C229 0.065 C230 0.060 C233 0.072C235 0.090 C236 0.088 C237 0.363 C238 0.104 C239 0.107 C242 0.744 P2X2/3Compound No. IC50 (μM) C2 0.2490 C3 0.3411 C4 0.1583 C9 0.3788 ReferenceCompound C10 1.8200 C12 0.0567 C13 0.5992 C15 0.7930 C20 0.0757 C210.200 C22 0.057 C23 0.6440 C33 0.262 C39 0.243 C43 0.1800 C44 0.4228 C520.589 C61 0.592 C65 0.506 C66 0.154 C71 0.067 C72 0.089 C84 0.742 C850.205 C88 0.122 C91 0.271 C92 0.222 C95 0.163 C111 0.129 C117 0.487 C1180.366 C119 0.505 C127 0.234 C133 0.770 C134 0.685 C135 0.488 C136 0.416C137 0.567 C142 0.549 C145 0.623 C148 0.113 C149 0.266 C150 0.232 C1510.961 C152 0.375 C153 0.096 C154 0.046 C157 0.978 C162 0.429 C164 0.071C165 0.338 C172 0.209 C173 0.259 C174 0.145 C175 0.141 C176 0.984 C1790.043 C180 0.220 C181 0.078 C192 0.069 C194 0.446 C196 0.652 C199 0.333C201 0.498 C202 0.166 C203 0.275 C204 0.056 C207 0.163 C208 0.1055 C2090.209 C210 0.399 C211 0.121 C212 0.123 C213 0.295 C214 0.072 C215 0.1942C216 0.065 C218 0.383 C222 0.040 C226 0.1547 C228 0.125 C229 0.113 C2300.106 C233 0.125 C235 0.145 C236 0.134 C237 0.781 C238 0.206 C239 0.131

According to the experimental data in the above table, compared withreference compound 10 (the group corresponding to V¹ in Formula (I) ofthe present invention is a carbon-containing group, and the groupcorresponding to V² is a nitrogen atom), the test compounds (the groupscorresponding to V¹ in Formula (I) are all nitrogen-containing groupsand the groups corresponding to V² are all carbon-containing groups)have significantly increased inhibitory activity on the P2X3 and P2X2/3receptors. The remaining compounds of the present invention also havesignificantly increased inhibitory activity on the P2X3 and P2X2/3receptors.

Various modifications of the invention in addition to those describedherein will become apparent to those skilled in the art from theforegoing description. Such modifications are intended to fall withinthe scope of the appended claims. Each reference, including all patents,applications, journal articles, books and any other disclosure, referredto herein is hereby incorporated by reference in its entirety.

1-9. (canceled)
 10. A method for the treatment of a disease mediated bythe P2X3 and/or P2X2/3 receptor antagonist, wherein the method comprisesadministering to a subject in need thereof an effective amount of acompound having the structure of Formula (I) or a pharmaceuticallyacceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide,isotopically labeled compound, metabolite or prodrug thereof:

wherein: L is selected from the group consisting of C(═O), CRR′, NR, O,S, S═O and S(═O)₂; V¹ is selected from the group consisting of N,

and NR; V² is selected from the group consisting of CR⁶ and C(═O);

represents either a single bond or a double bond, provided that when

is a single bond, V¹ is NR and V² is C(═O); R and R′ are eachindependently selected from the group consisting of H, halogen, C₁₋₆alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated or partially unsaturatedC₃₋₁₀ cyclic hydrocarbyl, saturated or partially unsaturated 3- to10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl andC₆₋₁₂ aralkyl, and at most 2 ring members in the cyclic hydrocarbyl andheterocyclyl are C(═O); R¹, R², R³ and R⁶ are each independentlyselected from the group consisting of H, halogen, —CN, —NH₂, —OH, —SH,—Se—R, —Si(R)₃, C₁₋₆ alkyl, C₂₋₆ alkenyl, C₂₋₆ alkynyl, saturated orpartially unsaturated C₃₋₁₀ cyclic hydrocarbyl, saturated or partiallyunsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to14-membered heteroaryl, C₆₋₁₂ aralkyl, C₁₋₆ haloalkyl, —C(═O)R^(a),—OC(═O)R^(a), —C(═O)OR^(a), —OR^(a), —SR^(a), —S(═O)R^(a), —S(═O)₂R^(a),—S(═O)₂NR^(a)R^(b), —S(═O)(═NR)R^(a), —NR^(a)R^(b), —C(═O)NR^(a)R^(b),—C(═S)NR^(a)R^(b), —C(═NR)NR^(a)R^(b), —NR^(a)—C(═O)R^(b),—NR^(a)—C(═O)OR^(b), —NR^(a)—S(═O)₂—R^(b), —NR^(a)—C(═O)—NR^(a)R^(b),—C₁₋₆ alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a), —C₁₋₆alkylene-C(═O)R, —C₁₋₆ alkenylene-OR^(a), —O—C₁₋₆ alkylene-NR^(a)R^(b)and —P(═O)R^(a)R^(b); R⁴ and R⁵ are each independently selected from thegroup consisting of H, —C(═O)OR^(a), —NR^(a)R^(b), —NR^(a)—C(═O)R^(b),—NR^(a)—C(═O)OR^(b), —C₁₋₆ alkylene-NR^(a)R^(b), —C₁₋₆ alkylene-OR^(a),—C₁₋₆ alkylene-O—C₁₋₆ alkylene-OR^(a), C₁₋₆ alkyl, C₁₋₆ haloalkyl, C₂₋₆alkenyl, C₂₋₆ alkynyl, saturated or partially unsaturated C₃₋₁₀ cyclichydrocarbyl, saturated or partially unsaturated 3- to 10-memberedheterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl and C₆₋₁₂aralkyl; alternatively, R¹ and R⁴ together form —NH—(C₁₋₆alkylene)-L-(C₁₋₆ alkylene)-, preferably —NHCH₂CH₂—O—CH₂CH₂—; the abovealkyl, alkylene, alkenyl, alkynyl, cyclic hydrocarbyl, heterocyclyl,aryl, heteroaryl and aralkyl, at each occurrence, are each optionallysubstituted with one or more substituents independently selected fromthe group consisting of halogen, hydroxyl, oxo, amino, cyano, nitro,—Si(R)₃, C₁₋₆ alkyl, saturated or partially unsaturated C₃₋₆ cyclichydrocarbyl, saturated or partially unsaturated 3- to 10-memberedheterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl, C₆₋₁₂ aralkyl,—C(═O)R^(a), —OC(═O)R^(a), —C(═O)OR^(a), —OR^(a), —SR^(a), —S(═O)R^(a),—S(═O)₂R^(a), —S(═O)₂NR^(a)R^(b), —NR^(a)R^(b), —C(═O)NR^(a)R^(b),—NR^(a)—C(═O)R^(b), —NR^(a)—C(═O)OR^(b), —NR^(a)—S(═O)₂—R^(b),—NR^(a)—C(═O)—NR^(a)R^(b), —C₁₋₆ alkylene-NR^(a)R^(b), —C₁₋₆alkylene-OR^(a), —C₁₋₆ alkenylene-OR^(a) and —O—C₁₋₆alkylene-NR^(a)R^(b), the alkyl, cyclic hydrocarbyl, heterocyclyl, aryl,heteroaryl and aralkyl are further optionally substituted with one ormore substituents independently selected from the group consisting ofhalogen, hydroxyl, oxo, amino, cyano, nitro, —NR^(a)R^(b), C₁₋₆ alkyl,—O—C₁₋₆ alkyl, saturated or partially unsaturated C₃₋₆ cyclichydrocarbyl, saturated or partially unsaturated 3- to 10-memberedheterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl and C₆₋₁₂aralkyl; and R^(a) and R^(b), at each occurrence, are each independentlyselected from the group consisting of H, —OH, C₁₋₆ alkyl, C₂₋₆ alkenyl,C₂₋₆ alkynyl, saturated or partially unsaturated C₃₋₁₀ cyclichydrocarbyl, saturated or partially unsaturated 3- to 10-memberedheterocyclyl, C₆₋₁₀ aryl, 5- to 14-membered heteroaryl and C₆₋₁₂aralkyl; alternatively, R^(a) and R^(b) together with the atom to whichthey are attached form a 3- to 12-membered heterocycle or heteroaromaticring, the above groups are further optionally substituted with one ormore substituents independently selected from the group consisting ofhalogen, hydroxyl, oxo, amino, cyano, nitro, C₁₋₆ alkyl, —O—C₁₋₆ alkyl,saturated or partially unsaturated C₃₋₆ cyclic hydrocarbyl, saturated orpartially unsaturated 3- to 10-membered heterocyclyl, C₆₋₁₀ aryl, 5- to14-membered heteroaryl and C₆₋₁₂ aralkyl.
 11. The method according toclaim 10, wherein R^(a) and R^(b), at each occurrence, are eachindependently selected from the group consisting of H, —OH, methyl,ethyl, n-propyl, isopropyl, cyclopropyl, phenyl, benzyl, methoxy andethoxy; alternatively, R^(a) and R^(b) together with the atom to whichthey are attached form a 5- to 8-membered heterocycle or heteroaromaticring.
 12. The method according to claim 10, wherein R¹, R², R³ and R⁶are each independently selected from the group consisting of H, F, Cl,Br, I, —CN, —NH₂, —OH, —SH, —Se—CH₃, —Si(CH₃)₃, —CH₂NH₂, —CH₂NHCH₃,—CH₂N(CH₃)₂, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl,tert-butyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, vinyl,propenyl, allyl, ethynyl, propynyl, trifluoromethyl, acetyl, —C(═O)OH,—C(═O)NH₂, —C(═S)NH₂, —C(═NH)NH₂, —NHCH₃, —NHCH₂CH₃, —NHCH₂CF₃,—N(CH₃)₂, —N(CH₃)(C₂H5), —N(C₂H5)₂, —NHCH₂CH₂OH, —NH—C(═O)CH₃,—NH—C(═O)CH═CH₂, methoxy, ethoxy, propoxy, phenyl, —NH—C(═O)—NH₂,—NH—C(═O)OCH₃, —SCH₃, —SCH₂CH₃, —SC(CH₃)₃, —SBn, —S(═O)CH₃, —S(═O)Bn,—S(═O)₂CH₃, —S(═O)₂Bn, —S(═O)₂NH₂, —S(═O)?NHCH₃, —S(═O)?N(CHR),—S(═O)(═NH)CH₃, —P(═O)(CHR), —P(═O)(C₂H₅)₂,


13. The method according to claim 10, wherein R⁴ and R⁵ are eachindependently selected from the group consisting of H, —C(═O)OC(CH₃)₃,—NH₂, —NHCH₃, —NHPh, —NHC(═O)CH₃, —NHBoc, methyl, ethyl, —CH₂CF₃,n-propyl isopropyl, n-butyl, isobutyl, tert-butyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, phenyl, benzyl,


14. The method according to claim 10, wherein the compound has thestructure of any of the following formulae:

preferably has the structure of any of the following formulae:


15. The method according to claim 10, wherein the compound has thefollowing structure:


16. A method for the treatment of a disease mediated by the P2X3 and/orP2X2/3 receptor antagonist, wherein the method comprises administeringto a subject in need thereof a pharmaceutical composition, thepharmaceutical composition comprises a prophylactically ortherapeutically effective amount of the compound according to claim 10,or a pharmaceutically acceptable salt, ester, stereoisomer, polymorph,solvate, N-oxide, isotopically labeled compound, metabolite or prodrugthereof, and a pharmaceutically acceptable carrier, and thepharmaceutical composition is preferably in the form of a solid,semi-solid, liquid, or gas preparation.
 17. The method according toclaim 10, wherein the disease is selected from the group consisting of aurinary tract disease selected from reduced bladder capacity, frequentmicturition, urge incontinence, stress incontinence, bladderhyperreactivity, benign prostatic hypertrophy, prostatitis, detrusorhyperreflexia, nocturia, urinary urgency, pelvic hypersensitivity,urethritis, pelvic pain syndrome, prostatodynia, cystitis, andidiopathic bladder hypersensitivity; pain disease selected frominflammatory pain, surgical pain, visceral pain, dental pain,premenstrual pain, central pain, pain due to burns, migraine and clusterheadaches, nerve injury, neuritis, neuralgia, poisoning, ischemicinjury, interstitial cystitis, cancer pain, viral, parasitic orbacterial infection, post-traumatic injury and pain associated withirritable bowel syndrome; cardiovascular system disease, preferablyhypertension; respiratory disease selected from chronic obstructivepulmonary disease, asthma and bronchospasm; gastrointestinal diseaseselected from irritable bowel syndrome (preferably diarrhea-dominantirritable bowel syndrome), inflammatory bowel disease, biliary colic,renal colic, and pain associated with gastrointestinal distension. 18.The method according to claim 16, wherein the disease is selected fromthe group consisting of a urinary tract disease selected from reducedbladder capacity, frequent micturition, urge incontinence, stressincontinence, bladder hyperreactivity, benign prostatic hypertrophy,prostatitis, detrusor hyperreflexia, nocturia, urinary urgency, pelvichypersensitivity, urethritis, pelvic pain syndrome, prostatodynia,cystitis, and idiopathic bladder hypersensitivity; pain disease selectedfrom inflammatory pain, surgical pain, visceral pain, dental pain,premenstrual pain, central pain, pain due to burns, migraine and clusterheadaches, nerve injury, neuritis, neuralgia, poisoning, ischemicinjury, interstitial cystitis, cancer pain, viral, parasitic orbacterial infection, post-traumatic injury and pain associated withirritable bowel syndrome; cardiovascular system disease, preferablyhypertension; respiratory disease selected from chronic obstructivepulmonary disease, asthma and bronchospasm; gastrointestinal diseaseselected from irritable bowel syndrome (preferably diarrhea-dominantirritable bowel syndrome), inflammatory bowel disease, biliary colic,renal colic, and pain associated with gastrointestinal distension.
 19. Amethod for the treatment of a disease mediated by the P2X3 and/or P2X2/3receptor antagonist, wherein the method comprises administering to asubject in need thereof a pharmaceutical composition, the pharmaceuticalcomposition comprises a prophylactically or therapeutically effectiveamount of the compound according to claim 15, or a pharmaceuticallyacceptable salt, ester, stereoisomer, polymorph, solvate, N-oxide,isotopically labeled compound, metabolite or prodrug thereof, and apharmaceutically acceptable carrier, and the pharmaceutical compositionis preferably in the form of a solid, semi-solid, liquid, or gaspreparation.
 20. The method according to claim 19, wherein the diseaseis selected from the group consisting of a urinary tract diseaseselected from reduced bladder capacity, frequent micturition, urgeincontinence, stress incontinence, bladder hyperreactivity, benignprostatic hypertrophy, prostatitis, detrusor hyperreflexia, nocturia,urinary urgency, pelvic hypersensitivity, urethritis, pelvic painsyndrome, prostatodynia, cystitis, and idiopathic bladderhypersensitivity; pain disease selected from inflammatory pain, surgicalpain, visceral pain, dental pain, premenstrual pain, central pain, paindue to burns, migraine and cluster headaches, nerve injury, neuritis,neuralgia, poisoning, ischemic injury, interstitial cystitis, cancerpain, viral, parasitic or bacterial infection, post-traumatic injury andpain associated with irritable bowel syndrome; cardiovascular systemdisease, preferably hypertension; respiratory disease selected fromchronic obstructive pulmonary disease, asthma and bronchospasm;gastrointestinal disease selected from irritable bowel syndrome(preferably diarrhea-dominant irritable bowel syndrome), inflammatorybowel disease, biliary colic, renal colic, and pain associated withgastrointestinal distension.